Magnetic solid phase extraction sorbents using methyl-parathion and quinalphos dual-template imprinted polymers coupled with GC-MS for class-selective extraction of twelve organophosphorus pesticides.

A novel magnetic dual-template molecularly imprinted polymer (DMIP) was prepared with methyl-parathion and quinalphos as templates. For comparison, a series of single-template polymers with only methyl-parathion (MPMIP) or quinalphos (QPMIP) as template as well as a non-imprinted polymer (NIP) in the absence of the template, were synthesized using the same procedure of DMIP. The obtained MIPs were characterized by scanning electron microscopy(SEM), Fourier transform infrared (FT-IR) spectroscopy, vibrating sample magnetometer (VSM), and X-ray diffraction (XRD). The properties including kinetic effect, thermodynamic effect, selectivity, and reusability of MIPs were investigated . Only DMIP possessed high affinity and good recognition for all twelve OPPs including quinalphos, isazophos, chlorpyrifos-methyl, chlorpyrifos, methidathion, triazophos, profenofos, fenthion, fenitrothion, methyl-parathion, parathion, and paraoxon in comparison to MPMIP, QPMIP, or NIP. Moreover, DMIP was used as magnetic solid phase extraction (MSPE) sorbent for the pre-concentration of twelve OPPs in cabbage samples. The developed DMIP-MSPE-GC-MS method showed high sensitivity, low LODs (1.62-13.9 ng/g), fast adsorption equilibrium (10 min), and acceptable spiked recoveries (81.5-113.4%) with relative standard deviations (RSD) in the range 0.05-7.0% (n = 3). The calibration plots were linear in the range 10-800 ng/mL with coefficients of determination (R2) better 0.99 for all twelve compounds. These results suggest that the DMIP is applicable for rapid determination and high throughput analysis of multi-pesticide residues. Graphical abstract.

Maize ZmMEK1 is a single-copy gene.

Mitogen-activated protein kinase (MAPK) cascade constitutes a conserved signaling module in eukaryotes. MAPK kinase (MAPKK) plays a crucial role in a MAPK cascade. ZmMEK1 is the first characterized MAPKK gene in maize. Although ZmMEK1 has been studied in detail in biochemical level, the genomic organization of ZmMEK1 gene is obscure. In this research, we clarified ZmMEK1 is a single-copy gene in the maize genome. Southern blot analysis using 3' specific region of ZmMEK1 cDNA as a probe revealed the presence of distinct single bands in each lane of EcoRI and HindIII. Although previous Southern blot analysis using full-length ZmMEK1 cDNA as a probe revealed several hybridizing bands, we showed here that all bands come from one genomic fragment corresponding to ZmMEK1 gene. Furthermore, ZmMEK1 was induced by PEG, abscisic acid (ABA), and salicylic acid (SA) and was down-regulated by NaCl.

Accuracy and efficiency of digitally fabricated all-ceramic crowns from conventional impressions and intraoral scans: A single-blind clinical randomized controlled trial.

PURPOSE: To evaluate the accuracy of intraoral scanners by comparing the marginal fit of 70 all-ceramic crowns fabricated from both conventional impressions and intraoral scans. MATERIALS AND METHODS: A total of 70 posterior teeth requiring single-crown restorations randomly underwent either intraoral scanning or conventional impression-taking followed by laboratory scanning of the casts in a parallel-group RCT. Subsequently, 70 monolithic all-ceramic crowns were CAD/CAM fabricated; only the impression technique differed. Marginal fit, internal fit, adjustment time required for insertion and occlusal contacts, and visual analog scale (VAS) scores assessing dentists' satisfaction with all of the crowns were clinically evaluated by a blinded and calibrated examiner. Data were analyzed using independent-samples t test and likelihood ratio test or Fisher exact test. All tests were performed with α = .05. RESULTS: The mean marginal fit with intraoral scanning (57.94 ± 22.51 µm) was better than with diagnostic cast scanning (82.98 ± 21.72 µm). The difference was statistically significant (P = .000). The differences in internal fit, adjustment time for crown insertion and occlusal contacts, and VAS scores were also significant, and the secondary outcomes were in favor of intraoral scanning. CONCLUSION: Within the limitations of this clinical trial, CAD/CAM-fabricated single-tooth restorations in the posterior region produced by an intraoral scanning technique using TRIOS was found to be a more accurate and efficient alternative to restorations based on conventional impressions in combination with the laboratory scanning technique.

A specific fluorescent probe for fast detection and cellular imaging of cysteine based on a water-soluble conjugated polymer combined with copper(II).

In pure water system, the specific and rapid detection of cysteine (Cys) is very important and challenging. Herein, a new optical probe was developed for the purpose based on the complex of cupric ion (Cu2+) with a water-soluble conjugated polymer, poly[3-(3-N,N-diacetateaminopropoxy)-4-methyl thiophene disodium salts] (PTCO2). The fluorescence of PTCO2 in 100% aqueous solution can almost completely extinguished by Cu2+ ions due to its intrinsic paramagnetic properties. Among various amino acids, only Cys causes immediately the efficient recovery of the Cu2+-quenched fluorescence of PTCO2 with ~31-folds fluorescence enhancement because of the stronger affinity of Cys to Cu2+ leading to the formation of Cu2+-Cys complex through Cu-S bond and separation of Cu2+ from weak-fluorescent PTCO2-Cu(II) ensemble and thereby restoring the free PTCO2 fluorescence. In tris-HCl buffer solution (2 mM, pH 7.4), the intensity of the restored fluorescence is linear with the concentration of Cys, ranging from 0 to 120 µM and the estimated detection limit of Cys is 3.3 × 10-7 M with the correlation coefficient R = 0.9981. In addition, the PTCO2-Cu(II) ensemble probe exhibits low cytotoxicity and good membrane penetration, and its application in living cell imaging of Cys has also been explored.

PEGylation of 6-amino-6-deoxy-curdlan for efficient in vivo siRNA delivery.

RNA interference (RNAi) is an evolutionarily conserved gene-silencing phenomenon that shows great promise for developing new therapies. However, the development of small interfering RNA (siRNA)-based therapies need to establish efficient delivery system that silences target genes with siRNA doses that is clinically feasible in humans. Here we report synthesis and in vivo study of a novel PEGylated curdlan-based nanoparticle, designated as 6AC-100PEG, obtained by conjugation of mPEG 2000 to 6-amino-6-deoxy-curdlan. The complex of siRNA/6AC-100PEG showed homogenous nanoparticles with an average diameter of 200nm. MTT assay indicated that 6AC-100PEG does not have apparent cytotoxicity. Systemic administration of a complex of siapoB/6AC-100PEG significantly reduced the level of apoB mRNA in mouse liver, indicating that 6AC-100PEG can efficiently deliver siRNA to mouse liver and induce RNAi. Administration of siRNA/6AC-100PEG to mouse did not elevate liver enzyme level in the serum, indicating that 6AC-100PEG nanoparticle is a promising in vivo siRNA delivery agent.

Metabolic flux analysis of Arthrobacter sp. CGMCC 3584 for cAMP production based on 13C tracer experiments and gas chromatography-mass spectrometry.

Arthrobacter sp. CGMCC 3584 are able to produce cAMP from glucose by the purine synthesis pathway via de novo or salvage biosynthesis. In order to gain an improved understanding of its metabolism, (13)C-labeling experiment and gas chromatography-mass spectrometry (GC-MS) analysis were employed to determine the metabolic network structure and estimate the intracellular fluxes. GC-MS analysis helps to reflect the activity of the intracellular pathways and reactions. The metabolic network mainly contains glycolytic and pentose phosphate pathways, the tricarboxylic acid cycle, and the inactive glyoxylate shunt. Hypoxanthine as a precursor of cAMP and sodium fluoride as an inhibitor of glycolysis were found to increase the cAMP production, as well as the flux through the PP pathway. The effects of adding hypoxanthine and sodium fluoride are discussed based on the enzyme assays and metabolic flux analysis. In conclusion, our results provide quantitative insights into how cells manipulate the metabolic network under different culture conditions and this may be of value in metabolic regulation for desirable production.

Secretory carrier membrane protein SCAMP2 and phosphatidylinositol 4,5-bisphosphate interactions in the regulation of dense core vesicle exocytosis.

Secretory carrier membrane protein 2 (SCAMP2) functions in late steps of membrane fusion in calcium-dependent granule exocytosis. A basic/hydrophobic peptide segment within SCAMP2 (SCAMP2 E: CWYRPIYKAFR) has been implicated in this function and shown to bind and sequester phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2 or PIP2] within membranes through an electrostatic mechanism. We now show that alanine substitution of tryptophan W2 within SCAMP2 E substantially weakens peptide binding to negatively charged liposomes; other substitutions for arginine R4 and lysine K8 have only limited effects on binding. Electron paramagnetic resonance analysis of liposomes containing spin-labeled PIP2 shows that R4 but not K8 is critical for SCAMP E binding to PIP2. The interfacial locations of SCAMP E and its structural variants within lipid bicelles measured by oxygen enhancement of nuclear relaxation are all similar. Corresponding point mutations within full-length SCAMP2 (SC2-R204A, SC2-K208A, and SC2-W202A) have been analyzed for biological effects on dense core vesicle exocytosis in neuroendocrine PC12 cells. With the same level of overexpression, SC2-R204A but not SC2-K208A inhibited secretion of cotransfected human growth hormone and of noradrenalin. Inhibition by SC2-R204A was the same as or greater than previously observed for SC2-W202A. Analysis of noradrenalin secretion by amperometry showed that inhibitory mutants of SCAMP2 decrease the probability of fusion pore opening and the stability of initially opened but not yet expanded fusion pores. The strong correlation between SCAMP2 E interactions with PIP2 and inhibition of exocytosis, particularly by SC2-R204A, led us to propose that SCAMP2 interaction with PIP2 within the membrane interface regulates fusion pore formation during exocytosis.

[Clinical application of IPS-empress 2 pressable all-ceramic crowns].

OBJECTIVE: To evaluate the clinical prosthetic effect of IPS-Empress 2 pressahie ceramic crowns. METHODS: 198 teeth of 70 patients were restored with IPS-Empress 2 pressahie ceramic crowns. The patients were asked to return in one week and every half year. The clinical prosthetic effect was evaluated. RESULTS: Through follow-up of 3-38 months, the veneer porcelain crowns of 3 teeth were broken. 2 crowns fall off due to teeth fracture, gingivitis occurred in 2 teeth, pulpitis or periapical periodontitis occurred in 3 teeth. The shades of 3 crowns were darkening. The prosthetic effect of 185 teeth was satisfied. The rate of satisfaction was 93.4%. CONCLUSION: IPS-Empress 2 pressable all-ceramic crown has the advantages of aesthetic effect, good hiocompatihility and simple fabrication. But its strength is not enough for posterior teeth and it can not cover the deep color of non-vital teeth and metal materials.