Comprehensive Profiling by Non-targeted Stable Isotope Tracing Capillary Electrophoresis-Mass Spectrometry: A New Tool Complementing Metabolomic Analyses of Polar Metabolites

Mass spectrometry (MS) driven metabolomics is a frequently used tool in various areas of life sciences; however, the analysis of polar metabolites is less commonly included. In general, metabolomic analyses lead to the detection of the total amount of all covered metabolites. This is currently a major limitation with respect to metabolites showing high turnover rates, but no changes in their concentration. Such metabolites and pathways could be crucial metabolic nodes (e.g., potential drug targets in cancer metabolism). A stable-isotope tracing capillary electrophoresis–mass spectrometry (CE-MS) metabolomic approach was developed to cover both polar metabolites and isotopologues in a non-targeted way. An in-house developed software enables high throughput processing of complex multidimensional data. The practicability is demonstrated analyzing [U-¹³C]-glucose exposed prostate cancer and non-cancer cells. This CE-MS-driven analytical strategy complements polar metabolite profiles through isotopologue labeling patterns, thereby improving not only the metabolomic coverage, but also the understanding of metabolism.     

Fundamental processes in long air gap discharges

The development of atmospheric lightning is initiated and sustained by the formation in virgin air of ‘streamer corona’ and ‘leader’ discharges, very similar to those observed in laboratory long sparks. Therefore, the experimental and theoretical investigations of these laboratory discharges have become of large interest to improve the physical knowledge of the lightning process and to develop self-consistent models that could be applied to new protection concepts.In the present paper the fundamental processes of the subsequent phases of long air gap discharges are analyzed, from the first corona inception and development to the leader channel formation and propagation. For all these processes simulations models are discussed that have been essentially derived and simplified by the authors, in order to develop sequential time-dependent simulation of the laboratory breakdown, with both positive and negative voltages. The possibility of extending these models to the case of natural lightning is discussed in the companion paper, presented in this same volume. To cite this article: I. Gallimberti et al., C. R. Physique 3 (2002) 1335–1359.     

Characterization of sulfate-reducing bacteria in yeast industry waste by microcalorimetry and PCR amplification

Sulfate-reducing bacteria (SRB) were isolated from anaerobic sludge of yeast factory wastewater treatment plant by cultivation on Postgate C medium. Microcalorimetry was used to monitor the anaerobic digestion processes and to measure the growth rates of sulfate-reducing bacteria. The maximum growth rates determined by microcalorimetry and ATP analysis were different—μ_max(dQ/dt) = 0.165 ± 0.008 h⁻¹ and μ_max(N_ATP) = 0.207 ± 0.013 h⁻¹. Experiments on the cultivation of SRB from yeast industry wastewater treatment plant in batch culture showed that during the first 20 h the concentration of sulfate decreased from 78.3 mM down to 62.2 mM while the increase of sulfide production was negligible. Perceptible amount of sulfide (7.82 mM) appeared on the 33.5 h of fermentation together with a peak on the power–time curve and considerable increase in the cell count (1.26 × 10⁹). First steps of sulfate metabolism (activation of sulfate by ATP sulfurylase, production of H₂) are accompanied by endothermic heat effects, therefore the values of thermal power remain moderate until the evolution of sulfide starts. The influence of green microalgae Chlorococcum sp. (preparation Biotreat 100) on the growth characteristics of microorganisms was also studied. Identification of one SRB strain was started by sequencing of PCR-amplified 16S rRNA gene. Two sets of primers were used for PCR amplification, both specific for domain Bacteria but giving different gene fragments. PCR-products were purified with JETQUICK kit according to the manufacturer instructions.     

Hexaphyrin(1.0.1.0.0.0). A new colorimetric actinide sensor

Hexaphyrin(1.0.1.0.0.0) (isoamethyrin) undergoes a significant color change in the presence of UO₂²⁺, PuO₂²⁺, and NpO₂²⁺. The complexation of the first of these dioxo actinide cations was studied in semi-quantitative fashion in 1:1 MeOH-CH₂Cl₂. Under these conditions, the detection limit for UO₂²⁺ was found to be ca. 5.8 ppm by naked eye monitoring and <28 ppb by UV-vis spectroscopy. Isoamethyrin does not undergo a color change in the presence of most transition metals or when exposed to Gd(III). Isoamethyrin thus constitutes an attractive alternative to 2,2′(1,8-dihydroxy-3,6-disulfonaphthylene-2,7-bisazo)-bisbenzenarsonic acid (AzIII) and 2-(5-bromo-2-pyridylazo)-5-(diethylamino)phenol (BrPADAP), systems currently used as actinide cation sensors.     

Experimental design and the optimization of a polymer supported palladium complex for use in the Heck reaction

Optimization of the Heck reaction of 4-bromoacetophenone with styrene by a polymer supported, sulfur-containing palladacycle, varying 6 factors at a total of 28 different levels, corresponding to 5760 different possibilities was undertaken. Conversion improved from 34%, with large observable leaching to 88% with no leaching. This was accomplished using a Design of Experiments approach facilitated by the Statistical Design Package, MODDE 7.0^TM.     

Standardization of BSE rapid test performances and experiences gathered during the implementation of large-scale testing

The accumulation of pathological prion protein is used as a diagnostic marker for transmissible spongiform encephalopathies. According to European Union (EU) regulations cattle older than 30 months of age (Germany, France, Italy, and Spain by national law >24 months) and slaughtered for human consumption must be tested by using rapid tests for bovine spongiform encephalopathy (BSE). Likewise fallen stock and clinically affected animals must be tested. This article gives a short overview of the incidence of BSE in Europe. The diagnostic hierarchy, i.e., the officially approved methodology for the confirmation of suspect rapid test cases, and the organization of the numerous laboratories involved in this large-scale testing for BSE are described. Special emphasis is given to necessary quality control measures currently in place for BSE rapid testing laboratories and to measures intended to assure a consistent performance of the commercially available rapid test kits.Presented at BERM-9—Ninth International Symposium on Biological and Environmental Reference Materials, 15–19 June 2003, Berlin, Germany.     

Cognitive Demands and Second-Language Learners: A Framework for Analyzing Mathematics Instructional Contexts

The issues involved in teaching English language learners mathematics while they are learning English pose many challenges for mathematics teachers and highlight the need to focus on language-processing issues related to teaching mathematical content. Two realistic-type problems from high-stakes tests are used to illustrate the complex interactions between culture, language, and mathematical learning. The analyses focus on aspects of the problems that potentially increase cognitive demands for second-language learners. An analytical framework is presented that is designed to enable mathematics teachers to identify critical elements in problems and the learning environment that contribute to increased cognitive demands for students of English as a second language. The framework is proposed as a cycle of teacher reflection that would extend a constructivist model of teaching to include broader linguistic, cultural, and cognitive processing issues of mathematics teaching, as well as enable teachers to develop more accurate mental models of student learning.