Nonnatural ganglioside and lysoganglioside lipoforms 2693 MATERIALS AND METHODS MaterialsAll chemicals were of analytical grade or the highest purity available. Water (H 2 O) used for buffers and solutions was purifi ed by an ultrapure H 2 O system (EASYpure UV/UF D8612, Werner Reinstwassersysteme, Leverkusen, Germany). Solvents used in reactions of oxygen-and moisture-sensitive compounds were in anhydrous form. 1-Propanol, methanol (MeOH), pyridine, and acetic anhydride were degassed before use. The native mixture of bovine brain gangliosides, Cronassial ® , which consists of 21% GM1, 40% GD1a, 16% GD1b, 19% GT1b, and 4% other gangliosides ( 25 ), was available in our lab. Sphingolipid ceramide N -deacylase (SCDase) and ␤ -galactosidase from bovine testes were from Sigma-Aldrich (Schnelldorf, Germany). Triton™ X-100, sodium taurodeoxycholate, Grubbs catalyst 2nd generation, HoveydaGrubbs catalyst 2nd generation, and Stewart-Grubbs catalyst were also from Sigma-Aldrich. For normal phase (NP) column chromatography, silica gel 60 (0.040-0.063 mm or 0.015-0.040 mm) from Merck (Darmstadt, Germany) was used. For desalting and reversed phase (RP) column chromatography, LiChroprep ® RP-18 (0.040-0.063 mm) from Merck was used. For TLC analysis, highperformance TLC (HPTLC) silica gel 60 F 254 and HPTLC silica gel 60 RP-18 plates from Merck were used. For anion-exchange chromatography, DEAE Sephadex™ A-25 from Amersham Pharmacia Biotech AB (Uppsala, Sweden) was used. Purifi cationDesalting was performed by the method of Wiliams and McCluer ( 26 ). The usage of polar eluents for column chromatography causes signifi cant contamination of column material in the products. We reduced this contamination to a minimum by the following method. NP silica gel columns of a very small column volume (approximately 1 ml column volume per 5 mg of product) were prepared in fi ltration columns from Supelco (Bellefonte, PA) using a mobile phase in which the substance is retained on the column. The substance was applied to the column and washed with three column volumes of the same solvent. Then, the substance was eluted by an appropriate solvent of higher polarity. C-NMR spectroscopy, Bruker Avance 300, 400, or 500 instruments (Billerica, MA) were used. For exchange of exchangeable protons by deuterium, gangliosides were dissolved in CD 3 OD and the solvent was removed in a stream of nitrogen. This process was repeated three times before measurement. For EI-MS, a MAT 95 XL sector fi eld instrument (Thermo Finnigan MAT GmbH, Bremen, Germany) or a MAT 90 sector fi eld instrument gangliosidosis ( 22 ). Both substances are potential biomarkers for these diseases ( 23 ), and the lysoganglioside standards prepared in this work can be used for their quantitative analysis. Analytical proceduresQuantifi cation of gangliosides by MS, including quantitative imaging MS, requires calibration substances. Hereby, calibration is defined as the establishment of a correlation between analyte concentration and mass spectrometer response. Three principle meth...

Implementation of the authors' IHE protocol, designed in close accordance with the DIVI (German Interdisciplinary Association of Intensive Care and Emergency Medicine) out-of-hospital emergency protocol, helps to collect important data in I-ES. Usage of the protocol resulted in better documentation of emergency situations in contrast to the information found in discharge letters alone. Nevertheless, documentation of I-ES still needs to be improved, especially concerning the completeness of records.

Using two examples, this paper argues for the benefits in properly examining the supposed 'normal' or 'standard' constructions in order to better understand certain aspects of innovation or even make extraordinary discoveries. The systematic examination of numerous roof trusses over single-nave churches in German-speaking Switzerland in the first example give us a vivid picture of the construction methods in various regions over a span of 250 years. Based on these constructions, one can see which systems or approaches in constructing roofs were carried forward and which were not. In the case of the late-medieval log buildings in central Switzerland it is shown that within a rather run-down residential building lies the potential of great discoveries. Through systematic surveys and with the help of dendrochronology, over 30 log buildings have been discovered and documented dating back as far as 1300.

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