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.     

Screening of biomarkers in rat urine using LC/electrospray ionization-MS and two-way data analysis

Biofluids, like urine, form very complex matrixes containing a large number of potential biomarkers, that is, changes of endogenous metabolites in response to xenobiotic exposure. This paper describes a fast and sensitive method of screening biomarkers in rat urine. Biomarkers for phospholipidosis, induced by an antidepressant drug, were studied. Urine samples from rats exposed to citalopram were analyzed using solid-phase extraction (SPE) and liquid chromatography mass spectrometry (LC/MS) analysis detecting negative ions. A fast iterative method, called Gentle, was used for the automatic curve resolution, and metabolic fingerprints were obtained. After peak alignment principal component analysis (PCA) was performed for pattern recognition, PCA loadings were studied as a means of discovering potential biomarkers. In this study a number of potential biomarkers of phospholipidosis in rats are discussed. They are reported by their retention time and base peak, as their identification is not within the scope of the study. In addition to the fact that it was possible to differentiate control samples from dosed samples, the data were very easy to interpret, and signals from xenobiotic-related substances were easily removed without affecting the endogenous compounds. The proposed method is a complement or an alternative to NMR for metabolomic applications.     

Nitrous oxide emission as affected by liming an acidic mineral soil used for arable agriculture

The effect of liming an acidic mineral soil (Dystric Nitosol from southern China), used for arable agriculture, on N₂O emission was studied in an incubation experiment. After the soil pH had been raised from pH 4.4 to 5.2, 6.7 and 8.1, soil samples were either amended with NH₄ ⁺ and incubated aerobically, favoring nitrification or, after application of NO₃ ^–, the incubation took place under anaerobic conditions, favoring denitrification. Gas sampling for N₂O determination and soil analyses were performed at regular intervals up to 13 days. Under nitrification conditions only small N₂O emission rates were observed (max. 6 g N kg^–1 d^–1) with significant differences between high and low pH values during the first 2 days of incubation. The nitrifying activity was low, even with high pH, and this, together with good aeration conditions, could partly explain the small N₂O evolution. During denitrification, however, cumulative N₂O emissions reached much higher values (1600 g N kg^–1 in comparison to 40 g N kg^–1 under nitrification conditions). N₂O emission during denitrification was significantly enhanced by increasing soil pH. Under alkaline conditions (pH 8.1) a large nitrite accumulation occurred, which was in line with the highest nitrate reductase activity determined in this treatment. The limited availability of organic carbon is probably the main reason for the absence of further reduction of NO₂ ^– to N₂O or N₂. At pH 6.7 the total N₂O emission was slightly higher than at pH 8.1, although the start of pronounced emissions was retarded and only small amounts of NO₂ ^– accumulated. Acid soil conditions caused either negligible (pH 4.4) or only small (pH 5.2) N₂O emissions. It can be concluded that these kinds of soil, used alternatively for production of upland crops or paddy rice, are prone to high N₂O emissions after flooding, particularly under neutral to alkaline conditions. In order to avoid major N₂O evolution and accumulation of nitrite, which can be leached into groundwater, the pH should not be raised to values above 5.5–6.     

Process conditions in X-ray lithography for the fabrication of devices with sub-micron feature sizes

This article describes the fabrication of polymer structures with lateral dimensions in the sub-micron regime using hard X-rays (λ_c ≈ 0.4 nm) from the electron storage ring ANKA. Spincoated polymethylmethacrylate (PMMA) grades have been analyzed with respect to development rates and contrast. The contrast has been determined to be constant over a wide dose regime but rapidly decreases for dose values below 1 kJ/cm³. Films with a thickness from 2 to 11 μm have been patterned using a high resolution X-ray mask consisting of 2 μm thick gold absorbers on a suspended 1 μm thick silicon nitride membrane. The fabrication of sub-micron X-ray lithography structures with feature sizes down to 400 nm is confined by the mechanical parameters of the resist material and the process conditions. Surface tension after development limits the achievable aspect ratio of isolated pillars and walls, depending on the actual resist height. PMMA structures have been successfully used as template for electroplating of 1 μm thick gold to demonstrate the fabrication capability of sub-micron scale metal parts.     

Experimental Modelling using Low Melting Point Metallic Melts: Relevance for Metallurgical Engineering

This paper describes model experiments with liquid metals of low melting point as an important tool to investigate the flow structure and related transport processes in melt flows being relevant for metallurgical applications. Three examples have been selected to demonstrate the benefit of such experiments, namely the electromagnetic stirring of a metallic melt in a pool, the directional solidification of Al‐Si alloys under the influence of an electromagnetically driven flow and the behaviour of a liquid metal bubble plume if a vertically travelling field is applied. Results of flow measurements are presented and the relevance for metallurgical processes is discussed.     

Managing the five tensions of IT-enabled decision support in turbulent and high-velocity environments

This chapter examines issues concerned with supporting managers and organizations with information and communication technologies as they decide and act in turbulent and high-velocity environments. In particular, the chapter identifies five key tensions in this context: (1) the tension between the need for quick decisions and the need for analytical decision processes; (2) the tension involving managerial need for action and the need for the safest execution of decisions that may be bold and risky; (3) the tension around empowering middle managers and management teams at various organizational levels in the midst of powerful and impatient top executives; (4) the tension between programmed, quick-action learning loops and the increased requirement for emergence and improvisation; and (5) the tension between expending effort to eliminate the digital divide with other organizations versus finding expedient ways to communicate through heterogeneous digital infrastructures. Each of the tensions, and how it can be managed, is illustrated through a case example. The chapter ends by suggesting that the management of these critical tensions needs to be fused into the day-to-day fabric of management practices for decision support processes. This article is part of the “Handbook on Decision Support Systems” edited by Frada Burstein and Clyde W. Holsapple (2008) Springer.     

Do infants segment words or recurring contiguous patterns?

Eight experiments tested the hypothesis that infants' word segmentation abilities are reducible to familiar sound-pattern parsing regardless of actual word boundaries. This hypothesis was disconfirmed. in experiments using the headturn preference procedure: 8.5-month-olds did not mis-segment a consonantvowel- consonant (CVQ word (e.g., dice) from passages containing the corresponding phonemic pattern across a word boundary (C#VC#; "cold ice"), but they segmented it when the word was really present ("roll dice"). However, they did not segment the real vowel-consonant (VC) word (ice in "cold ice") until 16 months. Yet, at that age, they still did not false alarm on the straddling CVC word. Thus, infants do not simply respond to recurring phonemic patterns. Instead, they are sensitive to both acoustic and allophonic cues to word boundaries. Moreover, there is a sizable developmental gap between consonant and vowel-initial word segmentation. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Characterization of the long time oxidation protection of fluorine implanted technical TiAl-alloys using ion beam methods

In the present work the oxidation resistance of fluorine treated technical TiAl-alloys was investigated. Single and double fluorine beam line implantation was found to improve the high temperature oxidation resistance of this class of materials with Al-contents higher than 40 at.%. Calculated and measured fluorine depth profiles were compared. It was shown that the alloying elements do not modify significantly the fluorine profile and do not disturb the halogen effect. After single and double fluorine implantation and for different oxidation stages (isothermal/thermocyclic conditions) the maximum of the fluorine profile was measured by PIGE (Proton Induced Gamma Emission). The fluorine maximum was found to be located at the metal/oxide interface. The time dependence of the fluorine profile was determined as well. Double implantation led to a slower growing alumina layer. In this case a F-reservoir is obtained and improves the long term oxidation resistance of TiAl-based alloys.