An improved PCR-based method for site directed mutagenesis using megaprimers

The antimicrobial activity of a combination of lactic acid and whey permeate fermented by a nisin-producing Lactococcus lactis strain was tested by the agar diffusion method using bacteria isolated from fish as test organisms. Lactic acid inhibited all bacterial strains studied, but nisin whey permeate inhibited Gram-positive bacteria only. The combination was more effective than lactic acid alone against Pseudomonas fluorescens and Staphylococcus hominis isolated from fish, and Pseudomonas aeruginosa ATCC9721 and Micrococcus luteus ATCC9341.     

Exposure to polycyclic aromatic hydrocarbons in occupational versus urban environmental air

OBJECTIVES: To evaluate the balance between occupational and environmental exposure to suspended particulate matter (SPM) and polycyclic aromatic hydrocarbons (PAHs), comparison measurements were performed in a coal-fired power plant and the urban atmosphere from the town nearby. METHODS: The analysis of SPM for PAH content was done according to a high-performance liquid chromatography (HPLC)-based method. The microscopic assessment was performed using scanning electron microscopy (SEM) by silver coverage of the samples derived by air filter. RESULTS: Contrary to expectations, the results showed low levels of particle-bound PAHs in the occupational environment (< 1 ng benzo(a)pyrene/m3 air) and high levels in urban air (range 80-1250 ng benzo(a)pyrene/m3). The SPM collected from the power plant exhibited non-respirable characteristics (particles larger than 10 microm), whereas urban SPM almost exclusively contained respirable airborne particles (<3 microm). CONCLUSIONS: The PAH burden, combined with the enhanced probability of respiratory absorption, confers a much greater hazard potential to the urban SPM. Under these conditions, in areas or countries in which old technologies remain in use, occupational exposure to SPM containing PAHs might represent a severe underestimation of the total risk as it does not take into account the background air pollution.     

Design of molecularly imprinted polymer grafts with embedded gold nanoparticles through the interfacial chemistry of aryl diazonium salts

A novel strategy is described for the preparation of highly sensitive molecularly imprinted (MIPs) sensors for dopamine. It combines mercaptobenzene diazonium salt as a coupling agent for immobilizing gold nanoparticles to gold electrodes and benzoyl benzene diazonium salt as photoinitiator of radical polymerization at the said gold nanoparticle-decorated gold electrodes. The MIP films were prepared by surface-initiated photopolymerization (SIPP) of methacrylic acid (MAA) as functional monomer (F) for dopamine (DA) the template molecule (T), and ethylene glycol dimethacrylate (EGDMA), the crosslinker (C). Dimethylaniline was employed as a hydrogen donor. The specificity and selectivity were demonstrated by square wave voltammetry (SWV). The detection limit was 0.35 nmol L^-1 (0.054 ng mL⁻¹). The sensor layers are stable and adherent to the surface through aryl layers. The originality and advantage of the process lie in the use of aryl diazonium salt as coupling agents for anchroring nanoparticles and MIP layers to the electrode surface in a simple and efficient way which ensures high sensing performance together with good surface-MIP adhesion. The same strategy can be extended to a broad range of templates.     

Thiuram Disulfides as Pseudo‐irreversible Inhibitors of Lymphoid Tyrosine Phosphatase

We screened a small library of thiuram disulfides for inhibition of lymphoid tyrosine phosphatase (LYP) activity. The parent thiuram disulfide, disulfiram, inhibited LYP activity in vitro and in Jurkat T cells, whereas diethyldithiocarbamate failed to inhibit LYP at the concentrations tested. Compound 13 , an N‐(2‐thioxothiazolidin‐4‐one) analogue, was found to be the most potent LYP inhibitor in this series, with an IC₅₀ value of 3 μM . Compound 13 inhibits LYP pseudo‐irreversibly, as evidenced by the time‐dependence of inhibition, with a K_i value of 1.1 μM and a k_inact value of 0.004 s^?1. The inhibition of LYP by compound 13 could not be reversed significantly by incubation with glutathione or by prolonged dialysis, but could be partially reversed by incubation with dithiothreitol. Compound 13 also inhibited LYP activity in Jurkat T cells.     

Substrate Selection Influences Molecular Recognition in a Screen for Lymphoid Tyrosine Phosphatase Inhibitors

Assay design is an important variable that influences the outcome of an inhibitor screen. Here, we have investigated the hypothesis that protein tyrosine phosphatase inhibitors with improved biological activity could be identified from a screen by using a biologically relevant peptide substrate, rather than traditional phosphotyrosine mimetic substrates. A 2000‐member library of drugs and drug‐like compounds was screened for inhibitors of lymphoid tyrosine phosphatase (LYP) by using both a peptide substrate (Ac‐ARLIEDNE‐pCAP‐TAREG‐NH₂, peptide 1) and a small‐molecule phosphotyrosine mimetic substrate (difluoromethyl umbelliferyl phosphate, DiFMUP). The results demonstrate that compounds that inhibited enzyme activity on the peptide substrate had greater biological activity than compounds that only inhibited enzyme activity on DiFMUP. Finally, epigallocatechin‐3,5‐digallate was identified as the most potent inhibitor of lymphoid tyrosine phosphatase activity to date, with an IC₅₀ of 50 nM and significant activity in T‐cells. Molecular docking simulations provided a first model for binding of this potent inhibitor to LYP; this will constitute the platform for ongoing lead optimization efforts.     

Hybrid Organic‐Inorganic Membranes on Porous Supports by Size Exclusion and Thermal Sintering of Fluorescent Polyphenylsilsesquioxane Nanoparticles

Hybrid organic–inorganic membranes were prepared by size‐exclusion deposition and thermal sintering of fluorescent polyphenylsilsesquioxane nanoparticles on a ceramic support. Fluorescent polyphenylsilsesquioxane particles were prepared by a two‐step co‐polymerization of phenyltriethoxysilane with 0.1 mol‐% N‐(triethoxysilylpropyl)dansylamide. Because they were larger than the pores in the support, the particles formed a layer on top of the ceramic support that was sintered at 300 °C into a homogeneous, non‐porous membrane. Fluorescence of the modified polyphenylsilsesquioxane provided a valuable tool to monitor particle deposition and assist in characterization of the membranes.

     

ION-SELECTIVE POLYMER-SUPPORTED REAGENTS

ABSTRACT

The design and synthesis of polymer-supported reagents that can selectively complex targeted metal ions from multi-component solutions will continue to be an important area of research into the 21^st century. Environmental remediation and sensor technology are only two of a number of areas in which such polymers can be applied. This paper reviews the recent literature with an emphasis on the key ligands that have been immobilized in order to better understand where this research is heading in the near future.     

Delignification and Bleaching Response of Earlywood and Latewood

Abstract

The delignification response in cooking and the impact of bleaching on earlywood and latewood were studied. Spruce earlywood and latewood chips were pulped by the kraft process and subsequently treated with one bleaching chemical at a time. In cooking, latewood required a higher alkali charge to reach the same kappa number. No difference in the light absorption coefficient between the different fiber types was observed. After oxygen delignification the earlywood fibers had a higher light absorption coefficient at the same kappa number. The difference in light absorbing material was maintained when bleaching was performed with chlorine dioxide, ozone, and peracetic acid. Hydrogen peroxide decreased the light absorbing structures in the earlywood to the same level as for latewood. The earlywood pulp had a higher brightness at a given kappa number than the latewood. The higher brightness remained through all bleaching operations and was primarily due to a higher light scattering ability.     

Separate Bleaching of Pulp Fractions Enriched in Earlywood and Latewood Fibers

Abstract

Unbleached softwood kraft pulp was fractionated in hydrocyclones into a fine fraction, enriched in earlywood fibers, and a coarse fraction, enriched in latewood fibers. The response to individual bleaching chemicals and the bleachability in bleaching sequences of the pulp fractions was studied.

The light absorption coefficient, light scattering coeffient, and brightness were higher in the unbleached fine fraction than in the coarse fraction. Hydrogen peroxide managed to reduce the light absorption coefficient of the earlywood enriched fine fraction more efficiently than that of the latewood enriched fraction.

In the TCF‐sequence the light absorption coefficient of the fine fraction was reduced to the level of the coarse fraction at a given consumption of bleaching chemicals. In the ECF‐sequence the difference in light absorption coefficient remained between the fractions. At a given consumption of bleaching chemicals, the fine fraction had higher brightness than the coarse fraction, 2%‐ and 1%‐units on the ISO‐scale in the TCF and ECF bleaching sequence, respectively.