Biochemical identification of transmembrane segments of the Ca(2+)-ATPase of sarcoplasmic reticulum

The transmembrane segments of sarcoplasmic reticulum Ca(2+)-ATPase were determined by trypsinization of cytoplasmic side-out intact sarcoplasmic reticulum vesicles. The membrane portion of tryptic digest comprising the transmembrane fragments, joined by the intravesicular segments, was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after labeling with fluorescein 5-maleimide in the presence of sodium dodecyl sulfate. In this way, seven fluorescent bands of tryptic fragments below 11 kDa were observed which were derived from 4 pairs of membrane spanning segments and one hydrophobic sequence at the C-terminal end. Two peptides of 10.8 and 10.6 kDa had the identical N-terminal sequence beginning at Glu826, representing the transmembrane segments M7 and M8 and their connecting loop. A band at 8.1 kDa contained one peptide beginning at Tyr36 (M1/loop/M2). A 7.7-kDa peptide starting at Leu253 (M3/loop/M4) and a 7.3-kDa peptide beginning at Ala752 (M5/loop/M6) were also observed. A band at 6.7 kDa contained two peptides, one beginning at Ser48 (M1/loop/M2) and another beginning at Tyr763 (M5/loop/M6). In addition, a 4-kDa peptide beginning at Met925 was observed. The size of this peptide did not allow for a complete pair of transmembrane segments, but this peptide could have been derived from trypsinolysis between the last pair of membrane spanning segments. These data therefore provide biochemical evidence for at least 8 transmembrane segments and perhaps two more at the C-terminal end of the enzyme.     

Mixed Mode VLSI Implementation of a Neural Associative Memory

A mixed mode digital/analog special purpose VLSI hardware implementation of an associative memory with neural architecture is presented. The memory concept is based on a matrix architecture with binary storage elements holding the connection weights. To enhance the processing speed analog circuit techniques are applied to implement the algorithm for the association. To keep the memory density as high as possible two design strategies are considered. First, the number of transistors per storage element is kept to a minimum. In this paper a circuit technique that uses a single 6-transistor cell for weight storage and analog signal processing is proposed. Second, the device precision has been chosen to a moderate level to save area as much as possible. Since device mismatch limits the performance of analog circuits, the impact of device precision on the circuit performance is explicitly discussed. It is shown that the device precision limits the number of rows activated in parallel. Since the input vector as well as the output vector are considered to be sparsely coded it is concluded, that even for large matrices the proposed circuit technique is appropriate and ultra large scale integration with a large number of connection weights is feasible.     

Simulation of the submerged energy nozzle-mold water model system using laser-optical and computational fluid dynamics methods

The present work describes quantitative digital particle image velocimetry measurements of a full-scale water model of a thin slab mold. Different casting speeds and two submerged entry nozzles with one and two outlet ports have been investigated. The flow pattern of the single-port nozzle shows a counterclockwise-rotating double vortex that is nearly steady-state but leads to high stationary surface waves. The flow jets out of the two-port nozzle oscillate and produce a transient flow pattern with low wave amplitudes. The amplitudes for the one-port nozzle show a linear variation with the volumetric flow rate. The experimental results lead to a good interpretation of the flow phenomena and are used to validate steady-state numerical simulations with the commercial program, CFX, on the basis of the Reynolds equations. To describe anisotropic turbulence effects, the Reynolds stress model (RSM) is used for the flat single-port nozzle and the standard k-ɛ model for the mold flow. The calculated mean velocities and wave amplitudes, predicted from pressure distribution at the water surface, are generally in the consensus of the experimental data. An erratum to this article is available at .     

Intramolecular crosslinking of poly(vinyl alcohol)

Poly(vinyl alcohol) is crosslinked in dilute solution (c=0.1 wt%) with glutaraldehyde. The reaction product is characterized by viscometry and gel permeation chromatography (g.p.c.). The intrinsic viscosity decreases with increasing degree of crosslinking and does not depend on temperature. G.p.c. reveals that the reaction product is not homogeneous, but consists of a mixture of particles with different sizes, possibly both intra- and intermolecularly crosslinked molecules. The intramolecularly crosslinked molecules are smaller in size than the initial polymer molecules and their size depends on the degree of crosslinking. They possess a narrow particle size distribution even if the initial polymer sample had a broad molecular weight distribution.     

Studies on the sample weight for ochratoxin A determination in raw coffee

Ochratoxin A is a mycotoxin that may cause damage to the kidneys and the immune system in man. Foods are examined for this contaminant by the laboratories responsible for official food control. At present, there are no specific provisions for the sampling and examination of ochratoxin A in foods. Therefore, recourse is made to the provisions for aflatoxins. They prescribe the processing of samples with a weight of up to 30 kg. The examination of raw coffee for ochratoxin A in conjunction with the German Food Monitoring Programme of 2000 was undertaken with various sample weights (5, 10 and 30 kg) in order to identify the influence of the sample weight on the result. Overall, the sample weight did not have an effect on the detection and level of the ochratoxin A content in the samples of raw coffee examined.     

Growth Stresses in the Oxide Scales on TiAl Alloys at 800 and 900°C

In the oxidation of TiAl alloys, the role of scale-growth stresses formed during oxidation has, thus far, been unknown. In the present paper the oxide-growth stresses were investigated by the deflection-test method in monofacial oxidation (DTMO) accompanied by acoustic-emission measurements. On unmodified surfaces the growth stresses are compressive and reach levels of around –100 MPa. At the same time, significant acoustic emission occurs indicating that even under isothermal conditions, stresses are relieved by a scale-cracking mechanism. For oxide scales on TiAl surfaces, which had been ion implanted with chlorine before oxidation, a very thin protective alumina layer is formed which, however, develops growth stresses in the range of several GPa, accompanied by intensive acoustic emission. In all stress–time curves, a dynamic situation is observed. This consists of phases of stress relief by scale microcracking and phases of stresses increase due to crack healing and further oxide growth. As a result, the level of stress as a function of oxidation time, is characterized by an oscillating course.     

Vom freiwilligen Seminar zur Q2-Pflichtveranstaltung

Definition of the problem: Ethical reasoning cannot be enforced. Voluntary understanding is a basic condition for the development of ethical competencies and attitudes. The aim of this survey was to investigate whether the presence of those students who attended our seminars compulsorily would have a negative effect upon the quality of the course. Methods and results: The participants of our small-group seminars in medical ethics were asked to complete a questionnaire on teaching aims, seminar atmosphere, moderation and importance of subjects. One hundred and ninety-two voluntary participants (2001/2) and 293 compulsory participants (2002/3) completed the questionnaire and were statistically compared. There was no significant difference between voluntary and compulsory participants in any of the important items. However, there were considerable differences in some items between male and female participants. Conclusion: We were surprised that there was no difference between the two groups. This leads us to hope that our concept of case-based small-group seminars will prove capable of realising our teaching aims for all students. The difference between male and female participants may have important didactic consequences and will be the subject of further studies.