Ultrasensitive proteomics using high-efficiency on-line micro-SPE-nanoLC-nanoESI MS and MS/MS

Ultrasensitive nanoscale proteomics approaches for characterizing proteins from complex proteomic samples of <50 ng of total mass are described. Protein identifications from 0.5 pg of whole proteome extracts were enabled by ultrahigh sensitivity (<75 zmol for individual proteins) achieved using high-efficiency (peak capacities of approximately 10(3)) 15-microm-i.d. capillary liquid chromatography separations (i.e., using nanoLC, approximately 20 nL/min mobile-phase flow rate at the optimal linear velocity of approximately 0.2 cm/s) coupled on-line with a micro-solid-phase sample extraction and a nanoscale electrospray ionization interface to a 11.4-T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer (MS). Proteome measurement coverage improved as sample size was increased from as little as 0.5 pg of sample. It was found that a 2.5-ng sample provided 14% coverage of all annotated open reading frames for the microorganism Deinococcus radiodurans, consistent with previous results for a specific culture condition. The estimated detection dynamic range for detected proteins was 10(5)-10(6). An improved accurate mass and LC elution time two-dimensional data analysis methodology, used to both speed and increase the confidence of peptide/protein identifications, enabled identification of 872 proteins/run from a single 3-h nanoLC/FTICR MS analysis. The low-zeptomole-level sensitivity provides a basis for extending proteomics studies to smaller cell populations and potentially to a single mammalian cell. Application with ion trap MS/MS instrumentation allowed protein identification from 50 pg (total mass) of proteomic samples (i.e., approximately 100 times larger than FTICR MS), corresponding to a sensitivity of approximately 7 amol for individual proteins. Compared with single-stage FTICR measurements, ion trap MS/MS provided a much lower proteome measurement coverage and dynamic range for a given analysis time and sample quantity.     

Hydrogen storage properties of MgH2 mechanically milled with α and β SiC

To understand the influence of various crystallographic phases on hydrogen storage properties, ball milling of MgH₂ with hexagonal (α) and cubic (β) SiC have been performed. Structural characterization of all samples has been done by X-ray diffraction (XRD) analysis, particle size analysis and scanning electron microscopy (SEM). Investigation of hydrogen desorption properties of prepared nanocomposites has been done using temperature programmed desorption (TPD) technique. Despite the results of structural and morphological characterization of obtained nanocomposites are very similar, TPD spectra show significant differences regarding existence of intermediate temperature peak. In the sample milled with hexagonal SiC this peak originates both from H₂ and H₂O, while in the sample milled with cubic phase it only comes from H₂O. Both samples exhibit low temperature H₂ peak at 385 K.     

Characterisation of black cumin,pomegranate and flaxseed meals as sources of phenolic acids

The article focuses on the extraction of ten phenolic acids from black cumin (Nigella sativa L.), pomegranate (Punica granatum L.) and flax (Linum usitatissimum L.) seed meals. The extracts have been fractionated as free, esterified and insoluble‐bound phenolic compounds and quantitatively determined by HPLC–PDA. The analysed meals can be utilised for obtaining valuable phenolic acids. However, the distribution of phenolic compounds varies depending on the meal source. The insoluble‐bound fraction has been the richest for the black cumin meal, both qualitatively and quantitatively, containing all ten analysed phenolics. In the pomegranate meal, the main phenolic has been gallic acid, accounting for nearly 48% in free form. The esterified form of the flaxseed meal has been abundant with ferulic (1025.44 ± 3.99 mg kg^?1 dry weight), caffeic and p‐coumaric acids. The total amount of phenolic acids would be underestimated if only free fractions would be taken into account, while neglecting esterified (for the pomegranate and flax meals) and insoluble‐bound fractions (for the black cumin and pomegranate meals).     

Temperature induced evolution of structure/microstructure parameters and their correlations with electric/magnetic properties of nanocrystalline Nickel ferrite

Nickel ferrite nanoparticles were annealed in order to find dependence of electric/magnetic properties on crystallite size. The following correlations of crystallite size with physical parameters were found: (a) lattice parameter decreases with the increase in size and it reaches value for bulk counterpart approximately for crystallites bigger than 7 nm, (b) ac electrical resistivity at room temperature increases with the increase in crystallite size, (c) for crystallites of ~7 nm or smaller electrical resistivity have maximum value at 50 °C, (d) the real part of permittivity at selected frequency generally decreases with the increase in crystallite size and (e) magnetization increases with the increase in crystallite size. Deviation of stoichiometry, cation polyvalence, and cation redistribution with annealing are the main factors that influence physical properties of Nickel ferrite nanoparticles.     

Discovery of New Classes of Compounds that Reactivate Acetylcholinesterase Inhibited by Organophosphates

Acetylcholinesterase (AChE) that has been covalently inhibited by organophosphate compounds (OPCs), such as nerve agents and pesticides, has traditionally been reactivated by using nucleophilic oximes. There is, however, a clearly recognized need for new classes of compounds with the ability to reactivate inhibited AChE with improved in vivo efficacy. Here we describe our discovery of new functional groups—Mannich phenols and general bases—that are capable of reactivating OPC‐inhibited AChE more efficiently than standard oximes and we describe the cooperative mechanism by which these functionalities are delivered to the active site. These discoveries, supported by preliminary in vivo results and crystallographic data, significantly broaden the available approaches for reactivation of AChE.     

Effects of Trichoderma harzianum on Photosynthetic Characteristics and Fruit Quality of Tomato Plants

The beneficial role of fungi from the Trichoderma genus and its secondary metabolites in promoting plant growth, uptake and use efficiency of macronutrients and oligo/micro-nutrients, activation of plant secondary metabolism and plant protection from diseases makes it interesting for application in environmentally friendly agriculture. However, the literature data on the effect of Trichoderma inoculation on tomato fruit quality is scarce. Commercially used tomato cultivars were chosen in combination with indigenous Trichodrema species previously characterized on molecular and biochemical level, to investigate the effect of Trichoderma on photosynthetic characteristics and fruit quality of plants grown in organic system of production. Examined cultivars differed in the majority of examined parameters. Response of cultivar Gružanski zlatni to Trichoderma application was more significant. As a consequence of increased epidermal flavonols and decreased chlorophyll, the nitrogen balance index in leaves has decreased, indicating a shift from primary to secondary metabolism. The quality of its fruit was altered in the sense of increased total flavonoids content, decreased starch, increased Bioaccumulation Index (BI) for Fe and Cr, and decreased BI for heavy metals Ni and Pb. Higher expression of swolenin gene in tomato roots of more responsive tomato cultivar indicates better root colonization, which correlates with observed positive effects of Trichodrema.     

Physical and stress-strain properties of wheat (Triticum aestivum) kernel

BACKGROUND: Two hard wheat varieties and one soft variety grown under the same agroecological conditions were analyzed for their physical and uniaxial stress–strain compression properties. RESULTS: The physical properties of wheat kernel were determined at a moisture content of 0.13 kg kg^?1 (wet basis), whereas the stress–strain compression test was conducted at a kernel moisture content from 0.082 to 0.433 kg kg^?1. Mean kernel lengths were 5.46 (5.37 and 5.38) mm, widths were 2.56 (2.47 and 2.62) mm and thicknesses were 2.12 (2.18 and 2.43) mm for Simonida, Dragana and NS 40S cultivars, respectively. Bulk densities were 791.34 (Simonida), 788.51 (Dragana) and 731.77 kg m^?3 (NS 40S). The force at the yield point was 241.46 N for Dragana (moisture content 0.133 kg kg^?1), 244.30 N for Simonida (0.136 kg kg^?1) and 164.90 N for NS 40S (0.433 kg kg^?1). CONCLUSION: The width and thickness of the analyzed kernels were small compared with the length, and bulk densities were also moderate. The yield point force values of the two hard varieties were 2.2 times higher than the values of the soft variety, at a moisture content of 0.136 kg kg^?1 for Simonida, 0.133 kg kg^?1 for Dragana and 0.141 kg kg^?1 for NS 40S. Copyright © 2011 Society of Chemical Industry     

Optimal cascade hydraulic control for a parallel robot platform by PSO

A new cascade load force control design for a parallel robot platform is proposed. A parameter search for a proposed cascade controller is difficult because there is no methodology to set the parameters and the search space is broad. A parameter search based on particle swarm optimization (PSO) is suggested to effectively search the parameters of the cascade controller. We used a unified mathematical model of a hydraulic actuator of parallel robot platform. These equations are readily applicable to various types of proportional valves, and they unify the cases of critical center, overlapped, and underlapped valves. These unified model equations are useful for nonlinear controller design. Simulation results show the advantages of the proposed optimal tuned cascade controller to solve the formulated tracking problem in relation to the classical proportional–integral (PI) controller.