In situ liquid-liquid extraction as a sample preparation method for matrix-assisted laser desorption/ionization MS analysis of polypeptide mixtures

A novel liquid-liquid extraction (LLE) procedure was investigated for preparation of peptide and protein samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). LLE using ethyl acetate as the water-immiscible organic solvent enabled segregation of hydrophobic and hydrophilic polypeptides in mixtures, thereby reducing the complexity of mass spectra obtained by MALDI MS. The LLE technique was optimized for rapid and sensitive in situ (on-target) sample preparation for MALDI MS analysis of proteins and peptides at low-picomole and subpicomole levels. Addition of MALDI matrix to the organic solvent enhanced the efficiency of the LLE-MALDI MS method for analysis of hydrophobic peptides and proteins. LLE-MALDI MS enabled the detection of the hydrophobic membrane protein bacteriorhodopsin as a component in a simple protein mixture. Peptide mixtures containing phosphorylated, glycosylated, or acylated peptides were successfully separated and analyzed by the in situ LLE-MALDI MS technique and demonstrate the potential of this method for enhanced separation and structural analysis of posttranslationally modified peptides in proteomics research.     

Proton transfer reaction mass spectrometry, a tool for on-line monitoring of acrylamide formation in the headspace of maillard reaction systems and processed food

The formation of acrylamide was measured in real time during thermal treatment (120-170 degrees C) of potato as well as in Maillard model systems composed of asparagine and reducing sugars, such as fructose and glucose. This was achieved by on-line monitoring of acrylamide released into the headspace of the samples using proton transfer reaction mass spectrometry (PTR-MS). Unambiguous identification of acrylamide by PTR-MS was accomplished by gas chromatography coupled simultaneously to electron-impact MS and PTR-MS. The PTR-MS ion signal at m/z 72 was shown to be exclusively due to protonated acrylamide obtained without fragmentation. In model Maillard systems, the formation of acrylamide from asparagine was favored with increasing temperature and preferably in the presence of fructose. Maximum signal intensities in the headspace were obtained after approximately 2 min at 170 degrees C, whereas 6-7 min was required at 150 degrees C. Similarly, the level of acrylamide released into the headspace during thermal treatment of potato was positively correlated to temperature.     

Separation of plutonium and neptunium species by capillary electrophoresis-inductively coupled plasma-mass spectrometry and application to natural groundwater samples

Capillary electrophoresis (CE) was coupled to ICPMS in order to combine the good performance of this separation technique with the high sensitivity of the ICPMS for the analysis of plutonium and neptunium oxidation states. The combination of a fused-silica capillary with a MicroMist AR 30-I-FM02 nebulizer and a Cinnabar small-volume cyclonic spray chamber yielded the best separation results. With this setup, it was possible to separate a model element mixture containing neptunium (NpO2(+)), uranium (UO2(2+)), lanthanum (La3+), and thorium (Th4+) in 1 M acetic acid. The same conditions were also suitable for the separation of various oxidation states of plutonium and neptunium in different aqueous samples. All separations were obtained within less than 15 min. A detection limit of 50 ppb identical with 2 x 10(-7) M (3-fold standard deviation of a blank) was achieved. To prove the negligible disturbance of the plutonium and neptunium redox equilibria during the CE separations, plutonium and neptunium speciation by CE-ICPMS in acidic solutions was compared with the results of UV/visible absorption spectroscopy and was found to be in good agreement. The CE-ICPMS system was also applied to study the reduction of Pu(VI) in a humic acid-containing groundwater at different pH values.     

Removal of Cd(II) and Pb(II) ions, from aqueous solutions, by adsorption onto sawdust of Pinus sylvestris

Fixation of heavy metal ions (Cd(II) and Pb(II)) onto sawdust of Pinus sylvestris is presented in this paper. Batch experiments were conducted to study the main parameters such as adsorbent concentration, initial adsorbate concentration, contact time, kinetic, pH solution, and stirring velocity on the sorption of Cd(II) and Pb(II) by sawdust of P. sylvestris. Kinetic aspects are studied in order to develop a model which can describe the process of adsorption on sawdust. The equilibrium of a solution between liquid and solid phases is described by Langmuir model. Scanning electronic microscopy (SEM) coupled with energy dispersive X-ray analysis (EDAX) and X-ray photoelectron spectroscopy (XPS) shows that the process is controlled by a porous diffusion with ion-exchange. The capacity of the metal ions to bind onto the biomass was 96% for Cd(II), and 98% for Pb(II). The sorption followed a pseudo-second-order kinetics. The adsorption of these heavy metals ions increased with the pH and reached a maximum at a 5.5 value. From these results, it can be concluded that the sawdust of P. sylvestris could be a good adsorbent for the metal ions coming from aqueous solutions. Moreover, this material could also be used for purification of water before rejection into the natural environment.     

Directional velocity estimation using focusing along the flow direction. I: Theory and simulation

A new method for directional velocity estimation is presented. The method uses beam formation along the flow direction to generate data in which the correct velocity magnitude can be directly estimated from the shift in position of the received consecutive signals. The shift is found by cross-correlating the beamformed lines. The approach can find the velocity in any direction, including transverse to the traditionally emitted ultrasound beam. The velocity estimation is studied through extensive simulations using Field II. A 128-element, 7-MHz linear array is used. A parabolic velocity profile with a peak velocity of 0.5 m/s is simulated for different beam-to-flow angles and for different emit foci. At 45/spl deg/ the relative standard deviation over the profile is 1.6% for a transmit focus at 40 mm. At 90/spl deg/ the approach gave a relative standard deviation of 6.6% with a transmit focus of 80 mm, when using 8 pulse-echo lines and stationary echo canceling. Pulsatile flow in the femoral artery was also simulated using Womersley's flow model. A purely transverse flow profile could be obtained with a relative standard deviation of less than 10% over the whole cardiac cycle using 8 pulse emissions for each imaging direction, which is sufficient to show clinically relevant transverse color flow images.     

Tissue reactions of subcutaneously implanted mixture of ε-caprolactone-lactide copolymer and tricalcium phosphate. An electron microscopic evaluation in sheep

Biodegradable polymers, mainly derivates of -hydroxy acids, are widely used today in oral- and maxillofacial surgery, orthopedics, and other fields of surgery. These biomaterials are well tolerated by living tissue and fracture fixation devices made of polylactic or polyglycolic acid are clinically widely used today. Still, there are some problems in application of biodegradable polymers. Abacterial inflammatory reactions have been noticed after the clinical introduction of these devices. Both swelling and pain at the site of implantation have also been reported. The etiology of this inflammatory reaction is still unknown, despite the numerous studies. Therefore, the aim of the present study was to further characterize this inflammatory reaction in detail, by electronmicroscopy. We prepared a mixture of -caprolactone–lactide copolymer and tricalcium phosphate and placed it in the dermis in 12 sheep. Follow-up times were 9, 14, 24, and 52 weeks. We found that the mixture caused a mild inflammatory reaction. There were no signs of cell damage. Fibroblasts, macrophages, and eosinophils were found adjacent to the copolymer. The mixture is easy to handle and can be moulded into different shapes in room temperature. The results encourage us to continue our studies to develop a filling material for bone defects.     

Co-development and composite clusters--the secular strength of Nordic telecommunications

During the 1980s and 1990s Finland and Sweden were on the internationalfrontier in telecommunications, pioneering the first-generationcellular system and leading in the development of the secondgeneration. This strength in telecommunications has developedunder various regulatory regimes in a complex industrial history,going back to the nineteenth century. To account for this Fenno-Swedishtelecom trajectory, the article starts out with Porter's modelof industrial competitiveness and theories of public procurement,and then focus the attention and analysis in two directions:(i) the historical role of advanced, research-intensive usersand competitive public–private development pairs; and(ii) the emergence and significance of a composite binationalclusters and their local agglomerations in the accelerated industrygrowth in the 1990s.     

Mechanical properties of young concrete: Part II: Determination of model parameters and test program proposals

The paper concerns testing and modelling of the mechanical properties required as input to calculation programs made for crack risk estimation of hardening concrete structures. The results from several test series on mechanical properties of young concrete as described in Part I of this paper, are further evaluated. Model parameters for the modified CEB 1990 Model Code-equations, are determined for six concrete mixes, all having a w/b-ratio on 0.40, for compressive strength, tensile strength and E-modulus. To make the models applicable for young concrete, at ₀-parameter is introduced to fix the time at which significant mechanical properties are present. A test program to determine the model parameters is proposed, based on the experience that compressive strength tests have the smallest statistical scatter and that they are simplest to carry out.