Two-dimensional map of Haemophilus influenzae following protein enrichment by heparin chromatography

Two-dimensional gel electrophoresis separates several hundred protein molecules in one single experiment and is efficiently used to study the products expressed by different genomes. Low-copy-number gene products are invisible on a stained two-dimensional map and must be enriched such that sufficient amounts are present for visualization and identification. We investigated the enrichment of proteins of the bacterium Haemophilus influenzae by chromatography on immobilized heparin which has affinity for growth and protein biosynthesis factors. Total soluble proteins of the microorganism were fractionated on Heparin-Actigel which resulted in enrichment of approximately 160 proteins. The eluates, representing about 40% of the applied proteins, were analyzed by two-dimensional gel electrophoresis and the protein spots were characterized by amino acid composition analysis and matrix-assisted laser desorption ionization mass spectrometry. The proteins enriched by chromatography on the heparin gel were not exclusively low-copy-number gene products and they did not exclusively belong to one single class of proteins. The proteins that bound to the heparin gel are indicated in a two-dimensional protein map which includes more than 110 newly identified proteins.     

Novel expression and regulation of gastrin gene in human ovarian cancer cell line, SW626

Gastrin-secreting tumors have been identified in ectopic locations including the ovary; the mechanisms regulating gastrin gene expression, its distribution, and signaling pathways in these ectopic tissues are not known. The purpose of our present study was to determine: (1) whether the gastrin gene and peptide could be detected in ovarian cancer cell lines, (2) if functional gastrin releasing peptide receptors (GRP-R) are present, and (3) whether gastrin gene expression is altered by GRP. Five ovarian cancer cell lines (SW626, OVCA 420, OVCA 429, OVCA 432, and OVCA 433) were analyzed. We identified gastrin gene and peptide expression in the SW626 cell line but not in the OVCA lines. SW626 cells express a functional GRP-R that is correctly coupled to the Ca2+ signaling pathway. Treatment of SW626 cells with bombesin, the amphibian equivalent of GRP, inhibited expression of the gastrin gene in a time- and dose-dependent fashion. The SW626 ovarian cancer cell line will provide a useful model to further define regulation and expression of both the gastrin gene and peptide in ectopic (nongastrointestinal) tissues.     

A comparative evaluation of new silicon carbide diodes and state-of-the-art silicon diodes for power electronic applications

Recent progress in silicon carbide (SiC) material has made it feasible to build power devices of reasonable current density. This paper presents results including a comparison with state-of-the-art silicon diodes. Switching losses for two silicon diodes (a fast diode, 600 V, 50 A, 60 ns Trr), an ultrafast silicon diode (600 V, 50 A, 23 ns Trr), and a 4H-SiC diode (600 V, 50 A) are compared. The effect of diode reverse recovery on the turn-on losses of a fast insulated gate bipolar transistor (IGBT) are studied both at room temperature and at 150 /spl deg/C. At room temperature, SiC diodes allow a reduction of IGBT turn-on losses by 25% compared to ultrafast silicon diodes and by 70% compared to fast silicon diodes. At 150 /spl deg/C junction temperature, SiC diodes allow turn-on loss reductions of 35% and 85% compared to ultrafast and fast silicon diodes, respectively. The silicon and SiC diodes are used in a boost converter with the IGBT to assess the overall effect of SiC diodes on the converter characteristics. Efficiency measurements at light load (100 W) and full load (500 W) are reported. Although SiC diodes exhibit very low switching losses, their high conduction losses due to the high forward drop dominate the overall losses, hence reducing the overall efficiency. Since this is an ongoing development, it is expected that future prototypes will have improved forward characteristics.     

Flavin adenine dinucleotide binding is the crucial step in alcohol oxidase assembly in the yeast Hansenula polymorpha

We have studied the role of flavin adenine dinucleotide (FAD) in the in vivo assembly of peroxisomal alcohol oxidase (AO) in the yeast Hansenula polymorpha. In previous studies, using a riboflavin (Rf) autotrophic mutant, an unequivocal judgement could not be made, since Rf-limitation led to a partial block of AO import in this mutant. This resulted in the accumulation of AO precursors in the cytosol where they remained separated from the putative peroxisomal AO assembly factors. In order to circumvent the peroxisomal membrane barrier, we have now studied AO assembly in a peroxisome-deficient/Rf-autotrophic double mutant (Δper1.rif1) of H. polymorpha. By sucrose density centrifugation and native gel electrophoresis, three conformations of AO were detected in crude extracts of Δper1.rif1 cells grown under Rf-limitation, namely active octameric AO and two inactive, monomeric forms. One of the latter forms lacked FAD; this form was barely detectable in extracts wild-type and Δper1 cells, but had accumulated in the cytosol of rif1 cells. The second form of monomeric AO contained FAD; this form was also present in Δper1 cells but absent/very low in wild-type and rif1 cells. In vivo only these FAD-containing monomers associate into the active, octameric protein. We conclude that in H. polymorpha FAD binding to the AO monomer is mediated by a yet unknown peroxisomal factor and represents the crucial and essential step to enable AO oligomerization; the actual octamerization and the eventual crystallization in peroxisomes most probably occurs spontaneously.     

Synthese von substituierten 5-Amino-1,2,4-triazolen

Synthesis of Substituted 5-Amino-1,2,4-triazoles N-Nitro-amidino-dithiocarbimidic acid dimethylester 1 reacts with monosubstituted hydrazines under formation of the substituted 5-amino-1, 2, 4-triazoles. Disubstituted hydrazines give under the same conditions N-nitro-amidino-S-methyl-isothiosemicarbazides. The reaction of some more hydrazine derivatives with 1 yields the adequate substituted 1, 2, 4-triazoles.     

Evaluation of the thermal performance of frame walls enhanced with paraffin and hydrated salt phase change materials using a dynamic wall simulator

The incorporation of phase change materials (PCMs) in building materials for use as latent heat storage and for potential reduction of energy requirements is an on-going field of study. In this paper, the development and testing of PCM-enhanced cellulose insulation for use in frame walls is presented. Two types of PCMs, paraffin-based and hydrated salt-based, were mixed into loose-fill cellulose insulation at concentrations of 10% and 20% (by the weight of the wallboard) in a 1.22 m × 1.22 m (48 in. × 48 in.) frame wall cavity. The thermally-enhanced frame walls were heated and allowed to cool down in a dynamic wall simulator that replicated the sun's exposure in a wall of a building on a typical summer day. Peak heat fluxes, total “daily” heat flows, and surface and air temperatures were measured and recorded. Results show that the paraffin-based PCM-enhanced insulation reduced the average peak heat flux by up to 9.2% and reduced the average total “daily” heat flow up to 1.2%. Because of the hydroscopic behavior of un-encapsulated hydrated salt, the hydrated salt-based PCM-enhanced insulation did not provide any thermal storage benefit.     

Detection probability of Campylobacter

A rapid presence/absence test for Campylobacter in chicken faeces is being evaluated to support the scheduling of highly contaminated broiler flocks as a measure to reduce public health risks [Nauta, M. J., & Havelaar, A. H. (2008). Risk-based standards for Campylobacter in the broiler meat chain. Food Control, 19, 372–381]. Although the presence/absence test is still under development, an example data set of test results is analysed to illustrate the benefit of the detection probability concept. The detection probability of Campylobacter increases with the logarithm of the Campylobacter concentration in faeces according to an S-shaped curve which stretches about 2–3 log units. The detection probability is 50% at a Campylobacter concentration of 7.4 × 10⁶ cfu/g. The uncertainty in the detection probability is 32% at the most for a 90% confidence interval. This type of information allows for realistic calculations on the Campylobacter status of different food processing paths after splitting. Usable quantitative estimates on detection probability await a data set of test results from a test that is ready for use or has similar properties.     

Laser-induced dynamic radiative corrections in the dressed-state and in the Floquet picture

We investigate QED field-dependent effects in radiative shifts of atomic levels driven by a strong laser field. Our focus is on indirect laser-induced corrections to the Lamb shift of an atomic reference state due to a strong laser field which near-resonantly couples two other states, but not the reference state. Our aim is to show how to generalize previous treatments of related scenarios to the case of an induced indirect coupling. We also derive an expression of self-energy corrections in the Floquet picture and analyse various coupling schemes with respect to the shift of Floquet quasi-energies. The semi-classical Floquet picture and the fully quantized dressed-state approach are contrasted.