Rh-Catalyzed Cascade C-H Activation/Annulation of N-Hydroxybenzamides and Propargylic Acetates for Modular Access to Isoquinolones.

A sequential Rh(III)-catalyzed C-H activation/annulation of N-hydroxybenzamides with propargylic acetates leading to the formation of NH-free isoquinolones is described. This reaction proceeds through a sequential C-H activation/alkyne insertion/intramolecular annulation/N-O bond cleavage procedure, affording a broad spectrum of products with diverse substituents in moderate-to-excellent yields. Notably, this protocol features the simultaneous formation of two new C-C/C-N bonds and one heterocycle in one pot with the release of water as the sole byproduct.

Nanobody-based electrochemical competitive immunosensor for the detection of AFB1 through AFB1-HCR as signal amplifier.

A novel nanobody (Nb)-based voltammetric immunosensor coupled with horseradish peroxidase concatemer-modified hybridization chain reaction (HRP-HCR) signal amplifying system is described to realize the rapid and ultrasensitive detection of AFB1. To design such an immunoassay, anti-AFB1 Nbs with smaller molecular size were coated densely onto the surface of Au nanoparticle-tungsten disulfide-multi-walled carbon nanotubes (AuNPs/WS2/MWCNTs) functional nanocomposites as an effective molecular recognition element, whereas AFB1-streptavidin (AFB1-SA) conjugates were ingeniously bound with biotinylated HCR dsDNA nanostructures as the competitor, amplifier, and signal report element. In the presence of AFB1 targets, a competitive immunoreaction was performed between the analyte and AFB1-SA-labeled HCR (AFB1-HCR) platform. Upon the addition of SA-modified polyHRP (SA-polyHRP), AFB1-HCR nanostructures containing abundant biotins were allowed to cross-link to a quantity of HRP by streptavidin-biotin chemistry for signal amplification and signal conversion. Under optimal conditions, the immunosensor displayed a good linear correlation toward AFB1 ranging from 0.5 to 10 ng mL-1 with a sensitivity of 2.7 µA • (mL ng-1) and an ultralow limit of detection (LOD) of 68 fg mL-1. The specificity test showed that the AFB1 immunosensor had no obvious cross-reaction with OTA, DON, ZEN, and FB1. The signal of this sensor decreased by 10.18% in 4 weeks indicating satisfactory stability, and its intra- and inter-laboratory reproducibility was 3.42~10.35% and 4.03%~12.11%, respectively. This biosensing system will open up new opportunities for the detection of AFB1 in food safety and environmental analysis and extend a wide range of applications in the analysis of other small molecules. Graphical abstract.

miR-338-3p confers 5-fluorouracil resistance in p53 mutant colon cancer cells by targeting the mammalian target of rapamycin.

Evidence demonstrate that p53 mutations and microRNAs (miRs) are important components of 5-FU resistance in colorectal cancer (CRC). miR-338-3p has been reported associated with cancer prognosis. However whether or not it influences chemotherapy sensitivity and the underlying mechanisms have not been elucidated. Here, three types of human colon cancer cell lines, HT29 (mutant p53), HCT116 (wild-type p53), and HCT116 p53-/- (deficient p53), were treated with 5-FU. We showed that expression of miR-338-3p was correlated with apoptosis and 5-FU resistance in colon cancer cells. Ectopic expression of miR-338-3p conferred resistance to 5-FU in HCT116 cells. Further experiments indicated that miR-338-3p mediated 5-FU resistance through down-regulation of mTOR expression. Moreover, inhibition of miR-338-3p in HT29 and HCT116 p53-/- cells increased their sensitivity to 5-FU treatment. Furthermore, we detected autophagy changes in our experiment because mTOR was known prominently regulating autophagy and the competition between autophagy and apoptosis in response to 5-FU was a mechanism influencing 5-FU sensitivity. Our results reveal a critical and novel role of miR-338-3p in the correlation of 5-FU resistance with p53 status. Moreover, the miR-338-3p inhibitor has the potential to overcome 5-FU resistance in p53 mutant colon cancer cells.

Rapid redox-response featured visual ascorbic acid sensor based on simple-assembled europium metal-organic framework.

A facile, fast and visible sensing platform for ascorbic acid (AA) detection has been developed based on self-assembled hydrangea-like europium metal-organic framework (HL-EuMOF). HL-EuMOF was synthesized through a simple one-step mixing process with Eu3+ and 1, 10-phenanthroline-2, 9-dicarboxylic acid at room temperature, which exhibited excellent properties including strong red fluorescence, long decay lifetime (548.623 µs) and good luminescent stability. Based on the specific redox reaction between Fe3+ and AA, the HL-EuMOF@Fe3+ was fabricated with "turn-off" response for AA, where the resulting Fe2+ displayed effective fluorescence quenching ability toward HL-EuMOF. The sensor demonstrated low detection limit (31.94 nM), rapid response time (30 s) and high selectivity. Integration of smartphone-assisted RGB analysis with HL-EuMOF@Fe3+ permitted convenient and visible quantitative determination of AA level. This approach also presented good detection performances in complex human serum and beverage samples, which could provide a valuable tool for AA detection in biomedical research and food industry.

AuNPs-BP-MWCNTs-COOH-based electrochemical immunosensor for the determination of deoxynivalenol in wheat flour.

Deoxynivalenol (DON) has drawn considerable attention for its obvious pathogenicity and wide use in agro-products, which cause a potential threat to human health. In this work, an electrochemical immunosensor is developed for the highly sensitive and selective detection of DON in wheat flour using AuNPs-BP-MWCNTs-COOH and antibodies. The AuNPs-BP-MWCNTs-COOH nanocomposite was prepared via an in situ reduction reaction and ultrasonic-assisted liquid-phase exfoliation. The nanocomposite exhibits a larger surface area, decent stability, excellent electron transfer capability, good protein binding capability and prominent specificity. The plentiful carboxyl group on the nanocomposite can bind to the amino group of the antibody, and AuNPs have an affinity for the sulfhydryl group of the antibody, which makes it feasible for the nanocomposite to load the antibody. The peak currents are plotted against the logarithm of DON concentration from 0.002 to 80 ng mL-1 with a limit of detection (LOD) of 0.5 pg mL-1. This approach establishes an effective label-free immunosensor platform for the detection of DON with high sensitivity and selectivity in various food and agricultural products.

Photocatalytic degradation of norfloxacin by magnetic molecularly imprinted polymers: influencing factors and mechanisms.

Novel magnetic molecularly imprinted polymers (MMIP) were prepared for selective removal of norfloxacin by effectively utilizing photocatalytic degradation and magnetic separation techniques. The imprinted material with titanium layer and multihole surface showed an excellent photocatalytic property. In this paper, the kinetics of photocatalytic degradation of norfloxacin by MMIP was explored, and the influences of environmental factors, including solution pH, humic acid, common ions and water media on photocatalytic performance of MMIP were elucidated. The results showed that MMIP had good adaptability and could degrade norfloxacin within 60 min, but the degradation rate constant decreased in surface water. Based on the identification of intermediate products, the possible degradation pathways of norfloxacin were analysed, speculating that it might be degraded into small molecules in the form of de-piperazine ring, de-carboxyl group and de-fluorine. Moreover, the mineralization ratio of norfloxacin could reach 84.2% after ultraviolet irradiation for 150 min, and the low cobalt release of MMIP enhanced the security of the material. The results of adsorption and degradation cycle tests showed that MMIP obtained by molecular imprinting technology had excellent performance in sustainable use for micro organic pollutants removal.

An Efficient HPLC-PDA Coupled With Supel™ Tox DON SPE Approach for the Analysis of Deoxynivalenol Contamination in Cereal Grains and Feedstuffs in Jiangxi Province.

Deoxynivalenol (DON) was commonly found in grains and feedstuffs, which can cause human chronic diseases. In this study, a quick and reliable method was developed for the determination of DON in grains and feedstuffs in Jiangxi Province market. The sample was extracted with acetonitrile-water (84:16, v/v), then purified by Supel™ Tox DON SPE column, and detected by high-performance liquid chromatography (HPLC). The results showed that the calibration curve of DON showed good linearity in the range of 0.01-10.0 µg/mL, and the correlation coefficient R2 = 0.9999. The recovery of DON in the spiked maize sample was 94.8-98.5% by spiking with DON at 0.2 µg/g, 1.0 µg/g, and 2.0 µg/g. The RSD was between 2.5 and 3.3%. This method was used to analyze 120 samples, including 90 grains and 30 feedstuffs, collected from the Jiangxi Province market. The results showed that 81 samples of grains were positive with 90.0% positive rates, and 30 samples of feedstuff were positive with 100% positive rates. Maximum concentration of DON was 0.7 µg/g in oat and 6.9 µg/g in wheat feedstuffs, respectively. Fortunately, the positive samples of grains were safe levels in comparison with National standards for food safety limits of mycotoxins in food (1.0 µg/g), while, the feedstuff of oats was over the Maximum Guideline Level of 16.7% (the Maximum Residue Limit, MRL is 5.0 µg/g). The results of this study about current DON pollution in the grains and feedstuffs on the Jiangxi Province market have not been previously reported.

Venlafaxine antagonizes the noradrenaline-promoted colon cancer progression by inhibiting the norepinephrine transporter.

Epidemiological studies have demonstrated that the use of antidepressants is associated with a decreased risk of colorectal cancer (CRC); however, the mechanisms behind this association are yet unknown. Adrenergic system contributes to the stress-related tumor progression, with norepinephrine (NE) mainly secreted from adrenergic nerve fibers. Norepinephrine serotonin reuptake inhibitors are successfully used antidepressants. This study demonstrates that a widely used antidepressant venlafaxine (VEN) antagonizes NE-promoted colon cancer in vivo and in vitro. Bioinformatic analysis suggested that NE transporter (NET, SLC6A2), a target of VEN, was closely associated with the prognosis of clinical patients with CRC. In addition, the knockdown of NET antagonized the effect of NE. The NET-protein phosphatase 2 scaffold subunit alpha/phosphorylated Akt/vascular endothelial growth factor pathway partially mediates the antagonizing effect of VEN on NE's actions in colon cancer cells. These were also confirmed by in vivo experiments. Our findings revealed for the first time that, in addition to its primary function as a transporter, NET also promotes NE-enhanced colon cancer cell proliferation, tumor angiogenesis, and tumor growth. This provides direct experimental and mechanistic evidence for the use of antidepressant VEN in the treatment of CRC and a therapeutic potential for repurposing existing drugs as an anti-cancer approach to improve the prognosis of patients with CRC.

Detection and electrocatalytic mechanism of zearalenone using nanohybrid sensor based on copper-based metal-organic framework/magnetic Fe3O4-graphene oxide modified electrode.

A novel and simple strategy was proposed for the determination of ZEA in breakfast cereal, maize powder and rice flour using an electrochemical nanohybrid sensor based on copper-based metal-organic framework (Cu-MOF)/magnetic Fe3O4-graphene oxide (Fe3O4-GO) modified electrode fabricated by the layer-by-layer assembled technique. The synthesized Cu-MOF with high porosity favorably improved the effective surface area and the analytical performance of nanohybrid sensing electrode. The crafted sensor has large surface area, high electron transfer, and satisfactory efficiency. ZEA was electrochemically detected in a wide linear range from 159.2 to 2865.2 ng mL-1 with LOD of 23.14 ng mL-1 under the optimal conditions. Moreover, the electrocatalytic mechanism of ZEA oxidation was proposed by density functional theory (DFT). A favorable energetic interaction was presented when Cu-MOF adsorbed on Fe3O4-GO, and a small new band appeared on the Fermi level energy (Ef) that facilitated the electron transfer between bands.

TDG suppresses the migration and invasion of human colon cancer cells via the DNMT3A/TIMP2 axis.

Background: Colorectal cancer (CRC) is one of the most common malignant tumors with high rates of recurrence and mortality. Thymine DNA glycosylase (TDG) is a key molecule in the base excision repair pathway. Recently, increasing attention has been paid to the role of TDG in tumor development. However, the specific functions of TDG in CRC remain unclear. Methods: The biological functions of TDG and DNA methyltransferase 3 alpha (DNMT3A) in CRC were evaluated using migration and invasion assays, respectively. A tumor metastasis assay was performed in nude mice to determine the in vivo role of TDG. The interaction between TDG and DNMT3A was determined via co-immunoprecipitation (Co-IP). Chromatin immunoprecipitation analysis (ChIP) was used to predict the DNA-binding site of DNMT3A. We also performed methylation-specific PCR (MSP) to detect changes in TIMP2 methylation. Results: TDG inhibited the migration and invasion of human colon cancer cells both in vitro and in vivo. TDG promoted the ubiquitination and degradation of DNMT3A by binding to it. Its interference with siDNMT3A also inhibits the migration and invasion of human colon cancer cells. Furthermore, the ChIP, MSP, and rescue experiments results confirmed that TDG accelerated the degradation of DNMT3A and significantly regulated the transcription and expression of TIMP2, thereby affecting the migration and invasion of human colon cancer cells. Conclusion: Our findings reveal that TDG inhibits the migration and invasion of human colon cancer cells through the DNMT3A-TIMP2 axis, which may be a potential therapeutic strategy for the development and treatment of CRC.

A highly-sensitive and selective antibody-like sensor based on molecularly imprinted poly(L-arginine) on COOH-MWCNTs for electrochemical recognition and detection of deoxynivalenol.

A new strategy to mimic antibody for electrochemical recognition and detection of deoxynivalenol (DON) using a highly-sensitive and selective antibody-like sensor based on molecularly imprinted poly(l-arginine) (P-Arg-MIP) on carboxylic acid functionalized carbon nanotubes (COOH-MWCNTs) was proposed. l-arginine as functional monomer was screened to prepare imprinted electrode via its electro-polymerization in the presence of DON onto the surface of COOH-MWCNTs electrode coupled with theoretical calculation. Surface morphology, structural characteristics, and electrochemical properties of P-Arg-MIP/COOH-MWCNTs were characterized by SEM, EDS, FTIR, and CV, respectively. P-Arg-MIP/COOH-MWCNTs displayed relatively high conductivity, high effective surface area, antibody-like molecular recognition and affinity, and a good response towards DON in a linear range from 0.1 to 70 µM with LOD of 0.07 µM in wheat flour samples with satisfactory recovery and feasible practicability in comparison with HPLC. This method provides a promising biomimetic sensing platform for the determination of mycotoxins in food and agro-products.

Norepinephrine transporter promotes the invasion of human colon cancer cells.

Epidemiological studies suggested the use of antidepressants to be associated with decreased risk of colorectal cancer (CRC). However, the underlying mechanism through which this decreased risk occurs remains elusive. The norepinephrine transporter (NET) is a target of antidepressants that maintains noradrenergic transmission homeostasis; however, little is known about its function in human CRC cells. The present study, using public datasets and immunohistochemistry approaches, revealed that NET was highly expressed in human CRC tissues with metastasis and in human colon cancer cells. Furthermore, knockdown of NET inhibited the invasive capability of human colon cancer cells. Additionally, epithelial (E)-cadherin expression was increased and Notch1 signaling was inhibited in NET-depleted colon cancer cells. These findings suggest that NET is highly expressed in human colon cancer, which is associated with the invasion of human colon cancer cells by influencing cell-cell adhesion through the Notch1-E-cadherin pathway. Thus, the present study revealed a novel function for NET and its downstream effectors in colon cancer cells, which will be valuable for future studies in a clinical setting.

Norepinephrine-CREB1-miR-373 axis promotes progression of colon cancer.

The adrenergic system contributes to the stress-induced onset and progression of cancer. Adrenergic fibers are the primary source of norepinephrine (NE). The underlying mechanisms involved in NE-induced colon cancer remain to be understood. In this study, we describe the function and regulatory network of NE in the progression of colon cancer. We demonstrate that NE-induced phosphorylation of cAMP response element-binding protein 1 (CREB1) promotes proliferation, migration, and invasion of human colon cancer cells. The downstream effector of NE, CREB1, bound to the promoter of miR-373 and transcriptionally activated its expression. miR-373 expression was shown to be necessary for NE-induced cell proliferation, invasion, and tumor growth. We confirmed that proliferation and invasion of colon cancer cells are regulated in vitro and in vivo by miR-373 through targeting of the tumor suppressors TIMP2 and APC. Our data suggest that NE promotes colon cancer cell proliferation and metastasis by activating the CREB1-miR-373 axis. The study of this novel signaling axis may provide mechanistic insights into the neural regulation of colon cancer and help in the design of future clinical studies on stress biology in colorectal cancer.

Zeolite Imidazolate Framework Membranes on Polymeric Substrates Modified with Poly(vinyl alcohol) and Alginate Composite Hydrogels.

Poly(vinyl alcohol)-sodium alginate composite hydrogels were first introduced to synthesize robust and well-intergrown zeolite imidazolate framework (ZIF)/polymer hollow fiber membranes. Through enough adsorption interaction with metal ions by chelation, sufficient nucleation sites for in situ metal-organic framework (MOF) preparation are provided. Using this method, we can not only easily prepare defect-free MOF membranes ignoring the complex modification process and seed deposition but also structurally fix crystalline MOF layers and greatly improve the stiffness and durability of MOF composite membranes. The strategy also gives the appropriate level of generality for synthesis of versatile dense MOF membranes on a variety of polymeric supports. The fabricated ZIF-8/polyethersulfone membrane presented remarkable gas separation performance with H2 permeance of as high as 9.66 × 10-7 mol m-2 s-1 Pa-1 and a high H2/CO2 separation factor of up to 29.0.

An Efficient Gas Chromatography-Mass Spectrometry Approach for the Simultaneous Analysis of Deoxynivalenol and Its Bacterial Metabolites 3-keto-DON and 3- epi-DON.

Deoxynivalenol (DON) is one of the major toxic secondary metabolites produced by Fusarium fungi in cereal grains. Among the many promising strategies of DON detoxification are the microbial and enzymatic ones, which transform DON to nontoxic DON metabolites. Thus, proper analytical methods are needed for those DON metabolites. In this study, a robust gas chromatography-mass spectrometry (GC-MS) procedure was developed and validated for the simultaneous analysis of DON and two of its bacterial metabolites, 3-keto-DON and 3- epi-DON. The procedure involves a straightforward vacuum drying and derivatization step before the subsequent GC-MS analysis. Following the optimized protocol, DON and these two metabolites were separated on a capillary column within 15 min. The linear ranges for the these compounds were 10 to 2,000 ng mL-1 with correlation coefficients >0.99. For DON, 3- epi-DON, and 3-keto-DON, the limits of detection were 0.8, 3.0, and 0.05 ng mL-1, and the limits of quantification were 2.6, 10.0, and 1.0 ng mL-1, respectively. For all three compounds, the obtained relative standard deviation was 1.2 to 5.5%, and the recovery rates were 89.5 to 103.6%. The developed method was further validated by analyzing DON metabolites resulting from the biotransformation of DON initiated by cell-free lysates of the bacterium Devosia mutans 17-2-E-8. The developed protocol was sensitive, precise, accurate, and robust for the determination of DON, 3- epi-DON, and 3-keto-DON in liquid media and potentially other complex matrices without interference from other compounds.

The enzymatic epimerization of deoxynivalenol by Devosia mutans proceeds through the formation of 3-keto-DON intermediate.

The enzymatic detoxification of deoxynivalenol (DON) is a promising mitigation strategy for addressing this mycotoxin contamination of cereal grains. A recently described bacterium, Devosia mutans 17-2-E-8, capable of transforming DON into its non-toxic stereoisomer 3-epi-DON, holds promise for the development of such applications. Earlier observations suggested that DON epimerization proceeds via a two-step catalysis with 3-keto-DON as an intermediate. The results of this study indicate that NADPH is required for DON epimerization by cell-free protein extracts of D. mutans, while high concentrations of glucose and sucrose have a suppressive effect. Chemically synthesized 3-keto-DON incubated with D. mutans protein fractions enriched by ammonium sulfate precipitation at 35-55% saturation selectively reduced 3-keto-DON to 3-epi-DON, but fell short of supporting the complete epimerization of DON. In addition, seven Devosia species investigated for DON epimerization were all able to reduce 3-keto-DON to 3-epi-DON, but only a few were capable of epimerizing DON. The above observations collectively confirm that the enzymes responsible for the oxidation of DON to 3-keto-DON are physically separate from those involved in 3-keto-DON reduction to 3-epi-DON. The enzymatic nature of DON epimerization suggests that the process could be used to develop genetically engineered crops or microorganisms, ultimately reducing foodborne exposure of consumers and farm animals to DON.

Preparation of molecularly imprinted polymer for use as SPE adsorbent for the simultaneous determination of five sulphonylurea herbicides by HPLC.

A high selective pre-treatment method for the analysis of sulphonylurea herbicides (SUHs) in rice grain samples based on molecularly imprinted solid-phase extraction (MISPE) was developed. The molecularly imprinted polymers (MIPs) were synthesised with high adsorption capacity and suitable particle size using pyrazosulphuron ethyl (PS) as the template and by screening three porogens and porogen volumes. The resulting MIPs showed high selectivity for PS. The MISPE procedure was developed for the purification and enrichment of PS, bensulphuron methyl (BSM), tribenuron methyl (TBM), metsulphuron methyl (MSM) and nicosulphuron (NS) from a rice grain sample prior to reversed-phase high-performance liquid chromatography (HPLC) analysis. Average recoveries of the PS, NS, MSM and BSM were 95.6%, 88.6%, 87.8% and 89.4%, respectively, but the recovery of TBM was lower (48.2%). This pre-treatment methodology for extracting SUHs was simple and cleaner extractions. The limits of detection ranged from 10.1 to 50.0ngL(-)(1). Five local rice grain samples were analysed by HPLC-MISPE, and PS was detected in all five samples, while BSM was detected in one sample.

Screening and identification of the antibacterial bioactive compounds from Lonicera japonica Thunb. leaves.

Our aim was to screen for antibacterial bioactive compounds from Lonicera japonica leaves. Staphylococcus aureus and Escherichia coli were used as the indicator bacteria. Bacteriostatic assay-guided extraction and stepwise partitioning of the samples yielded five compounds of interest. Antimicrobial activities of the compounds were determined using a disk diffusion assay. Extracts, fractions, and compounds from L. japonica leaves possessed considerable antibacterial activities against the tested bacterial strains and the most active fraction was attributed to J3B2, which primarily contained 3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid. Meanwhile, five bacteriostatic constituents were isolated (3-O-caffeoylquinic acid, secoxyloganin, luteoloside, 3,5-di-O-caffeoylquinic acid and 4,5-di-O-caffeoylquinic acid), among which, secoxyloganin was isolated for the first time from leaves. The antibacterial activity of the compounds was in the order of 3,5-bis-O-caffeoyl quinic acid, 4,5-bis-O-caffeoylquinic acid, luteoloside>3-O-caffeoylquinic acid>secoxyloganin. Our results suggested that the phenolic compounds might significantly contribute to antibacterial activity and were the most responsible for the bacteriostatic activity of L. japonica leaves.

Preparation of molecularly imprinted solid phase extraction using bensulfuron-methyl imprinted polymer and clean-up for the sulfonylurea-herbicides in soybean.

A pre-treatment methodology based on the molecularly imprinted solid phase extraction (MI-SPE) procedure was developed for the determination of bensulfuron-methyl (BSM), tribenuron-methyl (TBM), metsulfuron-methyl (MSM) and nicosulfuron (NS) in soybean samples. A molecular imprinted polymer (MIP) was prepared by precipitation polymerization using BSM as the template molecule, alpha-methacrylic acid (MAA) as the functional monomer, trimethylolpropane trimethacrylate (TRIM) as the cross-linker and dichloromethane as the porogen. The binding behaviors of the template BSM and its analogues on the MIP were evaluated by high performance liquid chromatography (HPLC). Then, solid phase extraction (SPE) with a BSM molecularly imprinted polymer (BSM-MIP) as adsorbent was investigated and the optimum loading, washing, and eluting conditions for MI-SPE of the selected BSM, MSM, TBM, and NS were established. The optimized MI-SPE procedure was used to extract the sulfonylureas and a high recovery was obtained in the soybean samples.