Potential effects of individual and combined exposure to tetraconazole and cadmium on zebrafish from the perspective of enantioselectivity and intestinal microbiota.

As the wide use of pesticides, they could form combined pollution with heavy metals, which would affect their environmental behaviors and toxic effects. Particularly, the effects would be more intricate for chiral pesticides. In this study, the accumulation and dissipation trends of tetraconazole enantiomers in zebrafish were investigated by individual and combined exposure of cadmium (Cd) and tetraconazole (including racemate and enantiomers) after confirming the absolute configuration of tetraconazole enantiomer. For the enantiomer treatments, Cd enhanced the accumulation of S-(+)-tetraconazole, but declined the concentrations of R-(-)-tetraconazole in zebrafish. The dissipation half-lives of tetraconazole enantiomers were extended by 1.65-1.44 times after the combined exposure of Cd and enantiomers. The community richness and diversity of intestinal microbiota were reduced in all treatments, and there were significant differences in R + Cd treatment. There was synergistic effect between Cd and S-(+)-tetraconazole for the effects on the relative abundances of Fusobacteria, Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. For R-(-)-tetraconazole, Cd mainly exhibited antagonistic effects. In the combined exposure of Cd and S-(+)-tetraconazole, the relative abundance changes of Cetobacterium (Fusobacteria, increase) and Edwardsiella (Proteobacteria, decrease) might affect the carbohydrate metabolism and energy metabolism, and led to the increase of S-(+)-tetraconazole bioaccumulation concentration. In the combined exposure of Cd and R-(-)-tetraconazole, Cd could increase the relative abundance of Edwardsiella (Proteobacteria), and affect the amino acid metabolism, which might reduce the bioaccumulation concentration of R-(-)-tetraconazole. This study reported for the first time that the abundance of intestinal microbiota in zebrafish might affect the bioaccumulation and dissipation of tetraconazole enantiomers, and would provide new insight for the study of combined pollutions.

Fabrication of poly-dopamine-modified magnetic nanomaterial and development of integrated QuEChERS method for 122 pesticides residue analysis in fruits.

A rapid and accurate integrated QuEChERS method was established for the determination of multi-pesticide residues in fruits. Poly-dopamine-modified magnetic nanomaterial (Fe3O4-pDA) was homemade and characterized. The prepared Fe3O4-pDA has the functional group of absorbing the saccharides, and can be used as co-adsorbent with 3-(N, n­diethyl amino) propyl trimethoxy-silane (PSA) in the developed integrated QuEChERS method to purify the fruit matrix, thus achieve the accurate determination of multi-pesticides residue. Grape was used as the representative sample to explore the influence of the salting out agent and each purification adsorbent on the pesticide recoveries. Under the optimized conditions, the proposed method showed good linearity for 92.6% of pesticides in the concentration range of 1-150 µg L-1 with method limit of quantitative (mLOQs) ranged from 10 to 18 µg kg-1. Spiked recoveries experiments were performed on four kinds of grapes and other fruits (apple, watermelon, pear, jujube and peach), in which satisfactory recoveries and precision were obtained for most of the pesticides. Meanwhile, comparison experiments also verified this method was superior to the traditional QuEChERS method in terms of convenient operation, high efficiency and low reagent consumption. The further real sample analysis was performed using this method, and the overall detection rate was 52%, while 2% of samples were exceeding the maximum residue limits. All results confirmed that the proposed method could be used for the rapid, simple, low-costing and effective analyses of trace multi-pesticides residue in fruit samples.

Fabrication of magnetic magnesium oxide cleanup adsorbent for high-throughput pesticides residue analysis coupled with supercritical fluid chromatography-tandem mass spectrometry.

A rapid and accurate analytical method was established for multiple pesticide residues in complex matrices based on magnetic dispersive solid phase extraction (d-SPE) and supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS). To develop an efficient magnetic d-SPE method, magnetic adsorbent modified with magnesium oxide (Fe3O4-MgO) was prepared via layer-by-layer modification and used as cleanup adsorbent for removal of interferences that contain a large number of hydroxyl or carboxyl groups in the complex matrix. The obtained Fe3O4-MgO coupled with 3-(N,N-Diethylamino)-propyltrimethoxysilane (PSA) and octadecyl (C18) were used as d-SPE purification adsorbents and their dosages were systematically optimized with Paeoniae radix alba as the matrix model. Combined with SFC-MS/MS, rapid and accurate determination of 126 pesticide residues in the complex matrix was achieved. Further systematic method validation showed good linearity, satisfactory recovery, and wide applicability. The average recoveries of the pesticides at 20, 50, 80, and 200 µg kg-1 were 110, 105, 108, and 109%, respectively. The proposed method was applied to complex medicinal and edible root plants, such as Puerariae lobate radix, Platycodonis radix, Polygonati odorati rhizoma, Glycyrrhizae radix, and Codonopsis radix. The average recoveries of the pesticides at 80 µg kg-1 in these matrices were 106, 106, 105, 103, and 105%, respectively with an average relative standard deviation range of 8.24-10.2%. The results demonstrated the feasibility and wide matrix applicability of the proposed method, which is promising for pesticide residue analysis in complex samples.

[QuEChERS-liquid chromatography-tandem mass spectrometry for determination of 22 triazole pesticide residues in Chinese herbal medicines].

Eight well-known herbals in Zhejiang Province, Zhebawei, are commonly used as traditional Chinese herbal medicines owing to their rich active ingredients. However, the unavoidable use of pesticides during agricultural production has led to pesticide residue problems in these herbs. In this study, a simple, rapid, and accurate method was established to determine 22 triazole pesticide residues in Zhebawei. An improved QuEChERS method was used for sample pretreatment, and Rhizoma Atractylodis Macrocephalae was used as a representative sample. The sample was extracted with acetonitrile to eliminate some polar and nonpolar compounds, pigments, and other impurities, and the purification effects of multiwalled carbon nanotubes (MWCNTs), amino-modified multiwalled carbon nanotubes (MWCNTs-NH2), carboxylated multiwalled carbon nanotubes (MWCNTs-COOH), crosslinked polyvinylpyrrolidone (PVPP), zirconium dioxide (ZrO2), 3-(N,N-diethylamino)-propyltrimethoxysilane (PSA), octadecyl (C18), and graphitized carbon black (GCB) were compared. MWCNTs-COOH and C18 were selected as the purification adsorbents, and their dosages were systematically optimized. The combination of 10 mg of MWCNTs-COOH and 20 mg of C18 was eventually selected as the purification adsorbents. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for analysis, and box graphs were plotted to present the dispersion of each group of recoveries, thus enabling the identification of the data outliers, dispersion distribution, and data symmetry. The established method was systematically verified and showed good linearity over the concentration range of 1-200 µg/L (except for bromuconazole, epoxiconazole, and etaconazole) with correlation coefficients >0.99. The average recoveries of the 22 pesticides at spiked levels of 10, 20, 100, and 200 µg/kg were in the range of 77.0%-115% with relative standard deviations (RSDs) <9.4%. The limits of detection and quantification were 1-2.5 µg/kg and 10-20 µg/kg, respectively. The applicability of the developed method to other herbals was investigated at 100 µg/kg, and the average recoveries of the target pesticides in different matrices ranged from 76.4% to 123% with RSDs <12.2%. Finally, the method established was used to detect triazole pesticide residues in 30 actual Zhebawei samples. The results showed that triazole pesticides were present in Bulbus Fritillariae Thunbergii and Dendranthema Morifolium. Difenoconazole was detected in Bulbus Fritillariae Thunbergii at contents ranging from 41.4 µg/kg to 110 µg/kg, while difenoconazole, myclobutanil, triadimenol and propiconazole were detected in Dendranthema Morifolium at contents ranging from 16.1 µg/kg to 250 µg/kg. The established method can meet the requirements for the accurate quantitative analysis of triazole fungicides in Zhebawei.

Green synthesis of Fe3O4@SiO2@Salg particles for organophosphorus pesticides removal: Mechanisms, biosafety, and application.

A biosafe magnetic Fe3O4@SiO2@Salg (GMS) was successfully developed via a simple and economic method. Fe3O4@SiO2 particles were selected as the magnetic core, and they directly interacted with sodium alginate, thus mitigating the use of cross-linking agents. The microstructure, properties, and formation mechanism of GMS were examined using FE-SEM, TEM, VSM, TGA, zeta potential, and FT-IR. The as-developed GMS was used as a desirable adsorbent to remove organophosphorus pesticides (OPPs) from water bodies, and the influence of different factors on the removal efficiencies of OPPs was investigated. Optimal preparation conditions were determined, and the excellent removal efficiencies of the 18 OPPs were achieved in this study (80.5%-100% for tap, river, and seawater). After the GMS was recycled eight times, the removal efficiencies of 18 OPPs were maintained at >80%. Adsorption kinetics and isotherm models showed that the adsorption process was accurately fitted with the Langmuir isotherm and pseudo-second-order models. The values of ΔG, ΔH, and ΔS were negative, indicating that the OPPs were adsorbed on the GMS spontaneously via an exothermal reaction, and chemisorption was the dominant adsorption mechanism. Overall, the as-developed GMS was biosafe as revealed by cytotoxicity experiments. GMS had good environmental security, stability, and reusability, and it exhibited excellent potential for removing OPPs from aqueous solutions without causing secondary pollution.

Fabrication and application of hydrangea-like magnetic titanium dioxide (Fe3O4-TiO2) particles: Development of magnetic "one-step" pretreatment method for multi-pesticide residues analysis in fish.

Magnetic titanium dioxide (Fe3O4-TiO2) was prepared and utilized as the cleanup adsorbent and separation medium to modify the QuEChERS method, affording a facile, robust, and rapid magnetic "one-step" pretreatment method for the determination of multi-pesticide residues in fish. The pretreatment key parameters, such as the dosages of the purification adsorbents (Fe3O4-TiO2 and PSA), the dehydrating and salting out reagents, were systematically optimized by the orthogonal test method. Under the optimal conditions, satisfactory results of method evaluation were obtained. Good linearity of 127 target analytes was obtained from 1 to 250 µg L-1. The recoveries of 127 analytes at five spiked levels of 10, 25, 50, 125, and 250 µg kg-1 ranged from 71.0% to 129% with RSDs less than 15.0%. The method LOQs (MLOQs) of 127 analytes were 10 µg kg-1, meeting the requirement for multi-pesticide residues analysis in fish. Additionally, this magnetic "one-step" method was used for the analysis of multi-pesticide residues in real fish samples collected from Zhejiang Province, China. In summary, this method can work as a viable tool for multi-pesticide residues monitoring in fish.

Additive manufactured polyether-ether-ketone composite scaffolds with hydroxyapatite filler and porous structure promoted the integration with soft tissue.

Additive Manufactured (AM) Polyether-ether-ketone (PEEK) orthopaedic implants offer new opportunities for bone substitutes. However, owing to its chemical inertness, the integration between PEEK implants and soft tissue represents a major challenge threatening the early success of the PEEK implants. Here we investigated the influence of hydroxyapatite (HA) fillers and porous structure of AM HA/PEEK scaffolds on the integration with soft tissue through in-vitro cellular experiments and in-vivo rabbit experiments. Among the animal experiments, HA/PEEK composite scaffolds with HA contents of 0, 20 wt%, 40 wt% and pore sizes of 0.8 mm, 1.6 mm were manufactured by fused filament fabrication. The results indicated that HA promoted the proliferation and adhesion of myofibroblasts on PEEK-based composites by releasing Ca2+ to active FAK and its downstream proteins, while the surface morphology of the scaffolds was also roughened by the HA particles, both of which led to the tighter adhesion between HA/PEEK scaffolds and soft tissue in-vivo. The macroscopic bonding force between soft tissue and scaffolds was dominated by the pore size of the scaffolds but was hardly affected by neither the HA content and nor the surface morphology. Scaffolds with larger pore size bonded more strongly to the soft tissue, and the maximum bonding force reached to 5.61 ± 2.55 N for 40 wt% HA/PEEK scaffolds with pore size of 1.6 mm, which was higher than that between natural bone and soft tissue of rabbits. Although the larger pore size and higher HA content of the PEEK-based scaffolds facilitated the bonding with the soft tissue, the consequent outcome of reduced mechanical properties has to be compromised in the design of the porous PEEK-based composite implants. The present study provides engineering-accessible synergistic strategies on material components and porous architecture of AM PEEK orthopaedic implants for improving the integration with soft tissue.

Development of rapid low temperature assistant modified QuEChERS method for simultaneous determination of 107 pesticides and relevant metabolites in animal lipid.

A modified QuEChERS LC-MS/MS method was developed for 107 pesticides analysis in animal lipids such as pork lard, mutton tallow, chicken oil and butter. The challenges for high fat matrices clean up were studied in details by optimizing dispersive purification adsorbents coupled with rapid low temperature assistant methods. The method validation was carried out using pork lard and further appplied to other matrices by testing their recoveries. Good linearities were obtained with correlation coefficients greater than 0.99. Sensitive LOQs ranged from 5.0-50.0 µg kg-1. Both inter-day and intra-day precisions were lower than 20% indicating the good precision and accuracy of this method. The method applied to four animal lipids with 93%∼100% of analytes revealed satisfactory recoveries (ranged from 70% to 120%) and RSD (≤20%) at 10 µg kg-1and 50 µg kg-1 spiking levels respectively related to the matrix.

Zeolitic imidazolate framework-8 as a dispersive solid phase extraction sorbent for simultaneous determination of 145 pesticide residues in polyphenol-rich plants.

Many plants have a high polyphenol content, which causes the matrix effect and makes the analysis of trace pesticide residues highly challenging. A common approach to improving matrix effects is to purify pesticides through the use of sorbents, but this requires a combination of multiple sorbents and extensive use. Zeolitic imidazolate framework-8 is widely used for pesticide analysis due to its high porosity, large specific surface area, and versatility. Here, we established and validated a modified quick, easy, cheap, effective, rugged, and safe method based on a zeolitic imidazolate framework-8 that was used to test the removal ability for polyphenols. To this end, 145 pesticide residues in peppermint, perilla, fennel, and mulberry leaves were analyzed by the modified method coupled with liquid chromatography with tandem mass spectrometry. The mean recoveries of all pesticide residues were in the range of 74.3-103.7%, with mean relative standard deviations ≤9.1% at spiked concentrations of 1, 10, 50, and 100 µg/kg for mulberry leaves. The limits of quantitation of the method ranged from 1 to 50 µg/kg. This study offers a reliable approach for the accurate quantitative analysis of various trace substances in the polyphenol-rich plants.

Low-dose cadmium affects the enantioselective bioaccumulation and dissipation of chiral penflufen in zebrafish (Danio rerio).

Pesticides are currently extensively used in agriculture, forestry, animal husbandry, and environmental hygiene, and their residues have become a global environmental problem, which can easily form combined pollution with heavy metals. The present study examined the effects of chronic (28 days) aqueous exposure of chiral penflufen (rac-penflufen, R-(-)-penflufen and S-(+)-penflufen), a widely used fungicide, with/without cadmium (Cd), a highly toxic heavy metal in zebrafish (Danio rerio). After rac-penflufen individual or combined exposure with Cd, the bioaccumulation and residual levels of S-(+)-penflufen were significantly higher than R-(-)-penflufen, and the effects of Cd were insignificantly. But for penflufen enantiomer, the effects of Cd were more serious for R-(-)-penflufen, which could increase the bioaccumulation (up to1.73 times), inhibit the dissipation (up to 32.3%) and enhance the residue (up to 5.35 times) of R-(-)-penflufen in zebrafish, decreasing the enantioselectivity. However, significant increase of S-(+)-penflufen concentrations was only found in viscera under co-exposure of Cd. The tissue distribution of penflufen enantiomers were not affected by the presence of Cd, and no interconversion of the two enantiomers occurred regardless of the presence of Cd. These findings indicated that co-contamination with Cd could increase the persistence of R-(-)-penflufen in zebrafish, thus increasing the environmental risks. The significant differences of Cd effects on chiral pesticide enantiomer and racemate indicated that the combined pollution of heavy metal and chiral pesticide might have enantiomer-specific, which should raise concern, and the enantioselective mechanism deserve further study.

Development of magnetic solid phase extraction using magnetic amphiphilic polymer for sensitive analysis of multi-pesticides residue in honey.

A sensitive and time-saving method for the determination of multi-pesticide residues in honey was developed using magnetic solid phase extraction (MSPE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Magnetic polymer (N-vinyl pyrrolidone-divinyl benzene) (MVP-DB) was fabricated and employed as the adsorbent for extraction and enrichment of multi-pesticide residues in honey. MVP-DB contains lipophilic benzene ring, divinyl group, and hydrophilic pyrrolidone group. The good hydrophilic and hydrophobic structure of MVP-DB not only ensures sufficient dispersion in honey samples, but also enhances the ability to enrich target analytes. The predominant factors affecting the recoveries of analytes were systematically investigated, affording a rapid and highly efficient MSPE method. Under the optimal conditions, the method was verified, including the recovery, precision, linearity, sensitivity, and matrix effects. The results displayed that these pesticides showed good linearity in the range of 2-250 µg L-1. The MLOQs were 0.5 µg kg-1. The recoveries of pesticides in honey at the pre-spiked concentrations of 0.5-25 µg kg-1 were 61.6%-112% with RSDs less than 18.2%. Hence, the developed method displayed good application prospect for the determination of multi-pesticide residues in honey samples.

Removal of Matrix Interferences by Nano-MgO and Co-Adsorbents for Accurate Multi-Pesticide Residue Analysis in the Chinese Medicinal Herb, Paeoniae Radix Alba.

A simple, accurate, and high-throughput analytical method was developed to detect 123 pesticide residues in Chinese medicinal herb Paeoniae Radix Alba (PRA) by introducing nano-MgO as a highly efficient purification material based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) design concept. Various PRA samples were extracted using 8 mL 0.5% acetic acid-acetonitrile solution and purified by a dispersive solid-phase extraction method with 30 mg nano-MgO, 40 mg primary secondary amine (PSA), and 40 mg octadecylsilane (C18) as the cleanup adsorbents, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). 70.7% of pesticides showed a weak matrix effect after the purification process, indicating that this method can give the precise quantitative analysis of trace pesticides residue. The method was systematically validated under optimal conditions in five different kinds of PRA samples; good linearity was observed in the concentration range of 0.5-250 µg/L or 1-250 µg/L. Pesticide recovery in each sample spiked at concentrations of 20, 50, and 200 µg/kg ranged from 98.0% to 111% and the mean relative standard deviation ranged from 2.72% to 5.70%. Furthermore, the method comparison with the traditional QuEChERS method suggested the feasibility, advantages, and potential application prospect of the present method for the multi-pesticide residue analysis in various PRA samples.

Low-dose cadmium stress increases the bioaccumulation and toxicity of dinotefuran enantiomers in zebrafish (Danio rerio)?

Co-occurrence of pesticides and heavy metals has attracted extensive attention. The enantioselective behaviors of dinotefuran to aquatic organisms have not been reported, and the effects of cadmium (Cd) was absent, which were investigated in this study at environmentally relevant concentrations. The enantioselective accumulation and elimination of dinotefuran enantiomers were observed in zebrafish, and it had tissue specificity. The S-dinotefuran concentrations were higher than R-dinotefuran in heads and viscera, but it was opposite in muscles. There existed competition between S-dinotefuran and R-dinotefuran, and the existence of S-dinotefuran might decrease the accumulation and elimination of the R-dinotefuran in zebrafish. When co-exposure to Cd and dinotefuran, the accumulation concentrations of dinotefuran enantiomers increased in zebrafish at the initial stage, which were opposite latterly. The accumulation concentrations of R-dinotefuran in R + Cd treatment in fish were 3.4 times higher than those in R-dinotefuran treatment, and the enantiomer fraction (EF) values changed from 0.484 to 0.195. The oxidative stress of S-dinotefuran on zebrafish was highest, followed by rac- and R-dinotefuran. Co-exposure to Cd led to toxicity increase for R-dinotefuran, the malonaldehyde (MDA) content decreased significantly in R + Cd treatment during 7-28 days, while obvious declination of MDA contents was found on the 28th day in R-dinotefuran treatment. Furthermore, compared to R-dinotefuran treatment, Cd increased the relative expression of cz-sod (3.4 times), cas3 (1.6 times) and p53 (5.7 times) in R + Cd treatment. The co-exposure of Cd might alter the environmental behaviors and toxicity effects of dinotefuran enantiomers in zebrafish, including the enantioselectivity. The effects of Cd on accumulation and toxicity of R-dinotefuran were greater than those on S-dinotefuran. Thus, it is necessary to consider the effects of coexistent metals to chiral pesticides in ecological risk. SUMMARIZES: The enantioselective accumulation and elimination of dinotefuran enantiomers had tissue specificity. Cd increased the accumulation and toxicity of R-dinotefuran in zebrafish.

Simultaneous determination of 114 pesticides in complex Chinese herbal medicine Fritillaria using ordered mesoporous carbon CMK-3 as a reversed-dispersive solid phase extraction sorbent.

Fritillaria, a traditional Chinese herbal medicine, is classified into many medicinal species and contains numerous complex components. It is thus difficult to simultaneously detect multiple pesticide residues in different Fritillaria species. An easy, reliable, and widely applicable analytical method based on a modified Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) method coupled with LC-MS/MS was developed to solve these problems encountered during pesticide residue analysis in complex Fritillaria matrices. Ordered mesoporous carbon CMK-3 and a primary secondary amine (PSA) were used as efficient purification sorbents by optimization of the QuEChERS process. Systematic method validation was performed for four species of Fritillaria. The matrix effect of pesticides varied among different Fritillaria species, and matrix-matched standard solutions were thus employed for quantitative analysis. The mean recoveries of all pesticides ranged from 88.6% to 95.5%, with mean relative standard deviations (RSD) lower than 6% at spiked concentrations of 30, 120, and 240 µg kg-1. The limits of quantification (LOQ) for the developed method were in the range of 30-120 µg kg-1. This method was further used to analyze 47 Fritillaria samples from Zhejiang province, China, and seven pesticides were detected in 22 Fritillaria samples. These results demonstrate that the developed method is suitable for an accurate analysis of multiple pesticide residues in various Fritillaria.

Magnetic covalent organic framework as a solid-phase extraction absorbent for sensitive determination of trace organophosphorus pesticides in fatty milk.

A simple and efficient magnetic solid-phase extraction (MSPE) method was established with magnetic covalent organic framework (COF) as adsorbent to enrich organophosphorus pesticides from fatty milk samples, followed by the sensitive determination via LC-MS/MS. The key parameters influencing the MSPE efficiency were comprehensively investigated to afford an optimized procedure. All the target analytes could be captured directly by magnetic COF from milk without protein precipitation, making the pretreatment rapid and convenient. Systematic method validation demonstrated its satisfactory linearity, recoveries (80.0-105 %), and precision (RSDs <12.3 %). The method limits of quantification were 0.2-0.5 µg L-1. A comparison experiment to the reported solid-phase extraction fully verified the present MSPE more rapid, accurate, and environment-friendly. Furthermore, FT-IR and XPS analysis were performed to reveal the adsorption mechanisms of magnetic COF to organophosphorus pesticides, which could offer guidance on the rational design of COF adsorbent for various target analytes.

Facile synthesis of a core-shell structured magnetic covalent organic framework for enrichment of organophosphorus pesticides in fruits.

A facile strategy was developed for the fabrication of a magnetic covalent organic framework (COF) via grafting of the monomers, 2,5-dihydroxyterephthalaldehyde (Dt) and 1,3,5-tris(4-aminophenyl) benzene (Tb) onto surface-modified Fe3O4 nanoparticles. The magnetic COF, named as magnetic COF-DtTb, was readily fabricated without high temperature or harsh reaction conditions. The synthesized magnetic COF-DtTb nanoparticles were fully characterized, presenting a regular core-shell spherical structure, large specific surface area, superparamagnetism, and good thermal stability. Their potential as an enrichment adsorbent was investigated to establish an efficient magnetic solid-phase extraction method for the determination of organophosphorus pesticide residues in fruits. Systematic method validation revealed good linearity in the concentration range of 1-200 µg L-1 (correlation coefficient >0.9957). The method limits of detection were in the range of 0.002-0.063 µg kg-1, the method limit of quantification was 1.00 µg kg-1 and recoveries ranged from 72.8% to 111% with RSDs lower than 12.3%. The results indicated that magnetic COF-DtTb possesses superior trace enrichment properties for organophosphorus pesticides in fruits.

Development, validation, comparison, and implementation of a highly efficient and effective method using magnetic solid-phase extraction with hydrophilic-lipophilic-balanced materials for LC-MS/MS analysis of pesticides in seawater.

To achieve multi-pesticides residue analysis in seawater, hydrophilic-lipophilic-balanced magnetic particles were designed and fabricated by swelling polymerization of divinyl benzene (DVB) and N-vinyl pyrrolidone (NVP) on the surface of Fe3O4@SiO2 magnetic particles. The ratio of DVB to NVP was adjusted to achieve a proper balance in hydrophilicity and lipophilicity. The obtained magnetic particles were systematically characterized by TEM, SEM, FT-IR and vibrating sample magnetization. Based on the optimized magnetic nanoparticles, a sensitive magnetic solid-phase extraction method was developed for the simultaneous pre-concentration and determination of 96-pesticide residues from large-volume seawater samples prior to being detected by liquid chromatography-tandem mass spectrometry. Recoveries of pesticides in spiked seawater samples (0.001, 0.01, 0.1, 1.0 µg L-1) ranged from 62% to 112% with RSDs less than 21%. The method limits of detection of 96 pesticides ranged from 0.13 to 0.42 ng L-1, the method limits of quantification of 96 pesticides ranged from 1.0 to 10 ng L-1. The method was successfully applied to pesticide residue analysis in water samples from Jiulong River Estuary of China, demonstrating the prospects of this technique as a potential method for the rapid determination of trace levels of multi-pesticide residues in seawater.

Bioactive Glass-Polycitrate Hybrid with Osteogenetic Ability Comparable to Autogenous Bone.

Significant progress in artificial bone grafts has been achieved in recent years. However, none of them has osteogenic ability that is close to autogenous bones. Hence, improving osteogenic ability of artificial bone is the most prominent and challenging task in this field. In addition, angiogenesis could provide a stable environment and nutrients for survival of the cells and also plays a crucial role to the success of bone transplantation. In the present study, we combined citrate polymer and bioactive glass together as hybrids at the molecular level (PEC-GS/BG), with the expectation of acquiring osteogenic ability and angiogenic ability to repair bone defect that could comparable to autogenous bones. In vitro and in vivo experiment on the femoral condyle critical defects model of Sprague-Dawley rats were conducted for a complete evaluation. In vivo, the bone mineral density (BMD) in defects was no significant difference between autogenous bone groups (517 ± 21 mg/cm³) and PEC-GS/BG groups (509 ± 21 mg/cm³) (p > 0 05) at 12 weeks post-surgery. The BMD of the femoral condyle in normal males at the same age was measured to be 557 ± 16 mg/cm³, only slightly higher than the above date, indicating a nearly complete repair of the defects. It was also found that PEC-GS/BG promoted angiogenesis due to it stimulated organism to release vascular endothecial growth factor (VEGF). PEC-GS/BG also showed great osteogenic ability that was close to autogeneous bones, but much better angiogenic ability. What's more, from both protein and cell levels, PEC-GS/BG accelerated differentiation and mineralization of MC3T3E-1 cells. Consequently, osteogenetic performance of PEC-GS/BG was almost same to autogenous bones in repairing bone defects. Considering the high demand in bone grafts and all the difficulties in autogeneous bone supply, the PEC-GS/BG hybrids developed in this study may open a new horizon for bone repair.

Neurotrophin-3 acts on the endothelial-mesenchymal transition of heterotopic ossification in rats.

Despite the fact that extensive studies have focused on heterotopic ossification (HO), its molecular mechanism remains unclear. The endothelial-mesenchymal transition (EndMT), which may be partially modulated by neuroendocrine cytokines is thought to play a major role in HO. Neurotrophin-3 (NT-3), which has neuroendocrine characteristics is believed to promote skeletal remodeling. Herein, we suggest that that NT-3 may promote HO formation through regulation of EndMT. Here, we used an in vivo model of HO and an in vitro model of EndMT induction to elucidate the effect and underlying mechanism of NT-3 on EndMT in HO. Our results showed that heterotopic bone and cartilage arose from EndMT and NT-3 promoted HO formation in vivo. Our in vitro results showed that NT-3 up-regulated mesenchymal markers (FSP-1, α-SMA and N-cadherin) and mesenchymal stem cell (MSC) markers (STRO-1, CD44 and CD90) and down-regulated endothelial markers (Tie-1, VE-cadherin and CD31). Moreover, NT-3 enhanced a chondrogenesis marker (Sox9) and osteogenesis markers (OCN and Runx2) via activation of EndMT. However, both EndMT specific inhibitor and tropomyosin-related kinase C (TrkC) specific inhibitor rescued NT-3-induced HO formation and EndMT induction in vivo and in vitro. In conclusion, our findings demonstrate that NT-3 promotes HO formation via modulation of EndMT both in vivo and in vitro, which offers a new potential target for the prevention and therapy of HO.

Locally Produced IGF-1 Promotes Hypertrophy of the Ligamentum Flavum via the mTORC1 Signaling Pathway.

BACKGROUND/AIMS: Narrowing of the lumbar spinal canal is a condition called lumbar spinal stenosis (LSS) and is a high-morbidity problem in the elderly. LSS is commonly caused by hypertrophy of the ligamentum flavum (HLF). Previous studies showed that fibrosis of the ligamentum flavum (LF) largely contributed to HLF. However, the underlying pathomechanism remains unclear. Insulin-like growth factor-1 (IGF-1) is known to have an intimate relationship with fibrosis in various tissues. Nevertheless, currently, there are few studies regarding IGF-1 in HLF. In this study, we investigated the role of IGF-1 in HLF and its potential molecular mechanism of action. METHODS: First, the IGF-1, phosphorylation of IGF-1 receptor (pIGF-1R), phosphorylation of AKT (pAKT), phosphorylation of S6(pS6), collagen I and collagen III expression levels were examined via immunohistochemistry and Western blotting in LF tissues from patients with LSS or Non-LSS. Second, primary LF cells were isolated from adults with a normal LF thickness and were cultured with different concentrations of IGF-1 with or without NVP-AEW541/rapamycin. RESULTS: The results showed that IGF-1, pIGF-1R, pAKT, pS6, collagen I and collagen III protein expression in the LSS group was significantly higher than that in the Non-LSS group. Meanwhile, pIGF-1R, pAKT, pS6, collagen I and collagen III protein expression was significantly enhanced in LF cells after IGF-1 exposure, which can be notably blocked by NVP-AEW541. In addition, pS6, collagen I and collagen III protein expression was blocked by rapamycin. CONCLUSIONS: Enhanced IGF-1 promotes the synthesis of collagen I and collagen III via the mTORC1 signaling pathway, which eventually contributes to hypertrophy of the ligamentum flavum.