Trennung von Muskelprotein, Kollagen und Elastin durch fraktionierte Extraktion mit Na-dodecylsulfat(SDS)-L?sungen

Zusammenfassung Die fraktionierte Extraktion mit Na-dodecylsulfat(SDS)-Lösungen ermöglicht die Trennung von Muskel- und Bindegewebs-Protein. In einem zweiten Schritt kann auch Kollagen von Elastin getrennt werden. Muskelprotein löst sich unter Schütteln bei Zimmertemperatur in einer SDS(2%)-ME(1%)-Lösung in einem Boratpuffer pH 9. Zur Lösung des Kollagens sind 100°C, intensives Rühren und eine SDS(8%)-ME(1%)-Lösung in einem Citratpuffer pH 5 notwendig. Das Verfahren wurde an nativem und hitzedenaturiertem Material angewendet. Die Möglichkeit zur Bestimmung des BEFFE-Wertes in Brühwürsten mit dieser Methode wird untersucht und die Anwendung auf andere Fleischprodukte diskutiert.

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Fractionation of muscle protein, collagen and elastin by extraction with buffers containing dodecylsulfat (SDS)

Summary Dodecylsulfate (SDS) containing buffers were used for stepwise extraction of proteins from muscle and connective tissue. The first step was done at room temperature with borate buffer pH 9 and the muscle proteins were extracted. The second treatment (citrate buffer pH 5 at 100°C) dissolved collagen and left elastin as residue. All extracts were analysed by electrophoresis to identify the proteins and evaluated by a biuret method insensitive to SDS and ME. This method was applied to native and heatdenatured proteins and to Brühwdrste. The possibility of applying this method to the determination of the BEFFE-content is discussed.

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Benutzte Abkürzungen NPN Nichteiweißstickstoffverbindungen - ATM Acetontrockenmasse - BEFFE bindegewebseiweißfreies Fleischeiweiß - OH-Pro Hydroxyprolin - SDS Natrium(Sodium)-dodecylsulfat - ME 2-Mercapto-äthanol - Tris Tris-(hydroxymethyl)aminomethan - PAGE Polyacrylamid-Gelelektrophorese - Vk Variationskoeffizient - S Standardabweichung - Mittelwert Die Arbeit ist Teil der Dissertation von I. de WreedeWir danken der Deutschen Forschungsgemeinschaft für ihre finanzielle Unterstützung und Herrn Dr. H. H. Heinert, Leiter des Staatl. Veterinär-Untersuchungsamtes Braunschweig, der freundlicherweise die Brühwurstproben zur Verfügung gestellt hat     

A proximate-time-optimal-control design and its application to a hard disk drive dual-stage actuator system

We propose a compensator-based strategy for design of a track-seeking and track-following control system for a dual-stage servo actuator in hard disk drives. A well-known decoupling structure is employed to disconnect the control of the primary voice coil motor (VCM) actuator from the loop for a secondary high-bandwidth actuator. The compensator is placed in the secondary loop and suitably combined with a saturation nonlinearity in order to obtain actuator signal boundedness. The design procedure consists of four steps: 1) design of an established nonlinear seek-settle-track following controller for the VCM; 2) design of a linear track following controller for the secondary actuator; 3) observer design; and 4) design of a compensator to retain global stability and to improve performance. The proposed control system improves performance of both long-span seeking (proximate-time-optimal controller) and short-span seeking. In addition, it achieves high-bandwidth track following performance. The experimental results show good track-following performance, and short-span/long-span-seeking performance with fast settling time. The overshoot during track seeking can be made negligible for a suitably tuned VCM-actuator control loop.     

Print your atomic force microscope

Progress in scanning probe microscopy profited from a flourishing multitude of new instrument designs, which lead to novel imaging modes and as a consequence to innovative microscopes. Often these designs were hampered by the restrictions, which conventional milling techniques impose. Modern rapid prototyping techniques, where layer by layer is added to the growing piece either by light driven polymerization or by three-dimensional printing techniques, overcome this constraint, allowing highly concave or even embedded and entangled structures. We have employed such a technique to manufacture an atomic force microscopy (AFM) head, and we compared its performance with a copy milled from aluminum. We tested both AFM heads for single molecule force spectroscopy applications and found little to no difference in the signal-to-noise ratio as well as in the thermal drift. The lower E modulus seems to be compensated by higher damping making this material well suited for low noise and low drift applications. Printing an AFM thus offers unparalleled freedom in the design and the rapid production of application-tailored custom instruments.     

The Small Molecule GAL-201 Efficiently Detoxifies Soluble Amyloid β Oligomers: New Approach towards Oral Disease-Modifying Treatment of Alzheimer’s Disease

Soluble amyloid β (Aβ) oligomers have been shown to be highly toxic to neurons and are considered to be a major cause of the neurodegeneration underlying Alzheimer’s disease (AD). That makes soluble Aβ oligomers a promising drug target. In addition to eliminating these toxic species from the patients’ brain with antibody-based drugs, a new class of drugs is emerging, namely Aβ aggregation inhibitors or modulators, which aim to stop the formation of toxic Aβ oligomers at the source. Here, pharmacological data of the novel Aβ aggregation modulator GAL-201 are presented. This small molecule (288.34 g/mol) exhibits high binding affinity to misfolded Aβ_1-42 monomers (K_D = 2.5 ± 0.6 nM). Pharmacokinetic studies in rats using brain microdialysis are supportive of its oral bioavailability. The Aβ oligomer detoxifying potential of GAL-201 has been demonstrated by means of single cell recordings in isolated hippocampal neurons (perforated patch experiments) as well as in vitro and in vivo extracellular monitoring of long-term potentiation (LTP, in rat transverse hippocampal slices), a cellular correlate for synaptic plasticity. Upon preincubation, GAL-201 efficiently prevented the detrimental effect on LTP mediated by Aβ_1-42 oligomers. Furthermore, the potential to completely reverse an already established neurotoxic process could also be demonstrated. Of particular note in this context is the self-propagating detoxification potential of GAL-201, leading to a neutralization of Aβ oligomer toxicity even if GAL-201 has been stepwise removed from the medium (serial dilution), likely due to prion-like conformational changes in Aβ_1-42 monomer aggregates (trigger effect). The authors conclude that the data presented strongly support the further development of GAL-201 as a novel, orally available AD treatment with potentially superior clinical profile.     

Microcavity-integrated graphene photodetector

There is an increasing interest in using graphene (1, 2) for optoelectronic applications. (3-19) However, because graphene is an inherently weak optical absorber (only ≈2.3% absorption), novel concepts need to be developed to increase the absorption and take full advantage of its unique optical properties. We demonstrate that by monolithically integrating graphene with a Fabry-Pérot microcavity, the optical absorption is 26-fold enhanced, reaching values >60%. We present a graphene-based microcavity photodetector with responsivity of 21 mA/W. Our approach can be applied to a variety of other graphene devices, such as electro-absorption modulators, variable optical attenuators, or light emitters, and provides a new route to graphene photonics with the potential for applications in communications, security, sensing and spectroscopy.     

Formal Translation of Bytecode into BoogiePL

Many modern program verifiers translate the program to be verified and its specification into a simple intermediate representation and then compute verification conditions on this representation. Using an intermediate language improves the interoperability of tools and facilitates the computation of small verification conditions. Even though the translation into an intermediate representation is critical for the soundness of a verifier, this step has not been formally verified. In this paper, we formalize the translation of a small subset of Java bytecode into an imperative intermediate language similar to BoogiePL. We prove soundness of the translation by showing that each bytecode method whose BoogiePL translation can be verified, can also be verified in a logic that operates directly on bytecode.